LOCUS       sequence-334721-        6203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  the host vector was obtained by substitution of the EGFP tag from a 
            pEGFP-N3-mGFAP plasmid for a HaloTag using BamHI and NotI 
            restriction sites.
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-Halo
FEATURES             Location/Qualifiers
     source          1..6203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1992..2882
                     /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    3007..3128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3044..3063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3098..3117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3135..3590)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3272..3293)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3484..3503
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3617..3721
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3723..4080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3723..3914
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3746..3766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3931..4066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4115..4909
                     /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             4115..4909
                     /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(4169..4188)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4779..4798
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5097..5116)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5141..5188
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5517..6105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6006..6025
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gcgctaccgg actcagatct cgagctcaag cttcgaattc aggcagggca
      661 ggatggagcg gagacgcatc acctctgcgc gccgctccta tgcctccgag acggtggtca
      721 ggggcctcgg tcctagtcga caactgggta ccatgccacg cttctccttg tctcgaatga
      781 ctcctccact ccctgccagg gtggacttct ccctggccgg ggcgctcaat gctggcttca
      841 aggagacacg ggcgagcgag cgtgcagaga tgatggagct caatgaccgc tttgctagct
      901 acatcgagaa ggtccgcttc ctggaacagc aaaacaaggc gctggcagct gaactgaacc
      961 agcttcgagc caaggagccc accaaactgg ctgatgtcta ccaggcggag cttcgggagc
     1021 tgcggctgcg gctggaccag cttacggcca acagtgcccg gctggaggtg gagagggaca
     1081 actttgcaca ggacctcggc accctgaggc agaagctcca agatgaaacc aacctgaggc
     1141 tggaggcaga gaacaacctg gctgcgtata gacaggaggc agatgaagcc accctggctc
     1201 gtgtggattt ggagagaaag gttgaatcgc tggaggagga gatccagttc ttaaggaaga
     1261 tctatgagga ggaagttcga gaactccggg agcagctggc ccaacagcag gtccacgtgg
     1321 agatggatgt ggccaagcca gacctcacag cggccctgag agagattcgc actcaatacg
     1381 aggcagtggc caccagtaac atgcaagaga cagaggagtg gtatcggtct aagtttgcag
     1441 acctcacaga cgctgcgtcc cgcaacgcag agctgctccg ccaagccaag cacgaagcta
     1501 acgactatcg ccgccaactg caggccttga cctgcgatct ggagtccctg cgcggcacga
     1561 acgagtccct agagcggcaa atgcgcgaac aggaagagcg ccatgcgcgg gagtcggcca
     1621 gttaccagga ggcacttgct cggctggagg aagagggcca aagcctcaag gaggagatgg
     1681 cccgccacct gcaggagtac caggatctac tcaacgttaa gctagccctg gacatcgaga
     1741 tcgccaccta caggaaattg ctggagggcg aagaaaaccg catcaccatt cctgtacaga
     1801 ctttctccaa cctccagatc cgagaaacca gcctggacac caaatccgtg tcagaaggcc
     1861 acctcaagag gaacatcgtg gtaaagactg tggagatgcg ggatggtgag gtcattaagg
     1921 actcgaagca ggagcacaag gacgtggtga tgcagaattc tgcagtcgac ggtaccgcgg
     1981 gcccgggatc catggcagaa atcggtactg gctttccatt cgacccccat tatgtggaag
     2041 tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc cctgtgctgt
     2101 tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg catgttgcac
     2161 cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac aaaccagacc
     2221 tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa gccctgggtc
     2281 tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc cactgggcca
     2341 agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc cctatcccga
     2401 cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc accaccgacg
     2461 tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg ccgatgggtg
     2521 tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc ctgaatcctg
     2581 ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt gagccagcga
     2641 acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct gtcccgaagc
     2701 tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct cgcctggcca
     2761 aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg ctgcaagaag
     2821 acaacccgga cctgatcggc agcgagatcg cgcgctggct gtcgacgctc gagatttccg
     2881 gctaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     2941 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3001 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3061 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3121 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3181 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3241 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3301 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3361 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3421 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3481 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3541 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3601 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3661 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3721 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3901 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4081 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4141 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4201 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4261 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4321 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4381 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4441 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4501 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4561 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4621 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4681 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4741 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4801 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4861 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4921 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4981 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5041 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5101 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5161 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5221 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5281 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5341 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5401 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5461 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5521 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5581 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5641 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5701 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5761 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5821 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5881 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5941 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6001 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6061 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6121 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6181 tcccctgatt ctgtggataa ccg
//
