LOCUS       sequence-141915-       10183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to drive expression in zebrafish astrocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10183)
  AUTHORS   Don EK, Formella I, Badrock AP, Hall TE, Morsch M, Hortle E, Hogan 
            A, Chow S, Gwee SS, Stoddart JJ, Nicholson G, Chung R, Cole NJ
  TITLE     A Tol2 Gateway-Compatible Toolbox for the Study of the Nervous 
            System and Neurodegenerative Disease.
  JOURNAL   Zebrafish. 2016 Sep 15.
  PUBMED    27631880
REFERENCE   2  (bases 1 to 10183)
  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: "Zebrafish. 
            2016 Sep 15."
COMMENT     Sequence Label: p5E-gfap
FEATURES             Location/Qualifiers
     source          1..10183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        690..708
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8157..8173)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8157..8173)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(8210..8228)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    8229..8352
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(8425..8443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8448..8464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8448..8464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8566..8585)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             8577..9386
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(8644..8663)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      9533..10121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10022..10041
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttggt aatacgactc actatagggc gaattgggta
      721 ccgggccccc cctcgaggta aggactgagg tgatgcacag tgttgggtat gttactttca
      781 ataagttatt ctttgctaat tattagtcat ttgaattgta ttgaaattac atttttagtt
      841 actttgccta aaaaaaaact tgataagtaa tcgaattaca tacttgaagt ccacataaat
      901 ctcattcatt cattccttct cttttcgact tagtcccttt attaatcagg ggtttctaca
      961 gcggaacgaa cctcatatgt ttcaatttag tgactatttt ttataaatta ttgacaatgt
     1021 ttttgttgct tgttcaaact acttatttaa aatgagctga aactgcacaa ttattgatat
     1081 ttcattggaa cagcttaatt ttttttatgt ttaatccact taaataaaat tccaattata
     1141 taatattcca attataaagt tacttaatca atttgtatta ggacaatatg aatgaattgt
     1201 gtggaaccct gcatttttta cagtgaacct ttccttgttt cttaaacatg ggtgattttc
     1261 ggagaaataa aatttgacaa tatctgaaat gtaaaaagaa aatattcagt tactctgcct
     1321 ttctaaatgc atagtttata ttatcatcag agttgtaaag taactagtta caaatactca
     1381 aattactgta attgagtagt ttttttcagg aattgtaatt tactaagtag ctaaaaaatg
     1441 tgtactttta ctttcccttg agtacattat taatgctgta acggtacttt tactccacta
     1501 ctttccttca gcctgcactc actactttgt tttatcttgt ctatgaggat tagaaaaatc
     1561 aatcctgtga ttccaatcaa atcgcacatc caaagtcttt acaaaagcat tttgtttgga
     1621 tgtgaaaatc gggttgatgt cagaactcaa tgtcagccca acgtcaatgt ccaacctaaa
     1681 atcaacctaa tatcaacgtc taatgatgtt acagcttgac gttgtgtggg cgttaccatt
     1741 atttgtcaga cattggattt tagttgccat acctgataaa taaatgtcag tatttgatgt
     1801 ccgtatgatc ttggatttag atgttggctc gacattggat tttggtcacc tgacataaca
     1861 acctaaatct atcctaataa taacgtctta tgacgttgtg tgccaaccat ttttgtttat
     1921 taaaaattta tataaacatt tacaataata tattatccat gtgatttaat agaaatgtta
     1981 tttgtttttt ttctccccca tatttctgaa tgacttaaag cttttattat caggataagg
     2041 ttgtggtttt tattttttgc ttgctaactt caattgctat tgcatttaca tttatgcaaa
     2101 tggggagacg cggtgggtca gttggcgttt ctgtgtggag tttgcatgtt ctccccgtgt
     2161 tcgcgtgggt tttctctggg tgctctggtt tcccctaaag acatgtagta taggtgaatt
     2221 gggtaagcta aattgtccgt agtgtgtatg tgtgaatgta tggatgtttt cctgtgatgg
     2281 gttgcagctg aaagggcatc cgctgtgtaa aacatatgct ggataagttg gcaattcatt
     2341 tcgctgtggt aaccccagat taatatcgga actacgccaa aaagaaaatg aatgaatgag
     2401 tgaattaatg caaatggctc atccttccac ctaattccag tgccttgcat tgcattcaag
     2461 ctctacaatt aatcagttcc tttattttca ctttccaaac tgaatccaac aactcatgag
     2521 gaccccaggg tgccttgatt taccagtgtc atgatgaaca caatctaccc cactttaacc
     2581 cactcgtcaa cataaatact cccttaaagg gtgtacttgt gtctgagtag aagtgtctgc
     2641 tgggttggag gtgtcgcaca ttccaggaca cgtcaagaag aacgctaaga gcaacaggag
     2701 aacaatgtca caaaaaaggt caaagaaacg ccatccatta gccagcttta taaacagctg
     2761 tctcgtggct aaagaagttc tcaagaataa ccaaccttgc atttttccac attagtttta
     2821 caccgcagca aagtcaccat gtggatttat agcatccaaa ccccattttc aatgagaggc
     2881 ttcacatttc tcgctctcat tcaatcccac actgcctctc ttttcagact aaatcctgat
     2941 aggatgagtg gtatgggggc ggggaggggg cacaatacct cagtccagct tccacaaaga
     3001 ctcaacacac ccatgcagat gtcctgttgt tggggaagtg catttgtaac caaatttcaa
     3061 actttcaact ttcatttgaa gcagtggaga ctgctccatt cctattcaag atcaatttat
     3121 tgaaatatga caacatttag aataactttg ttatatgtta ttatgtttta taatgcaatt
     3181 ttgtttctgt gacatacatt tttttattta gtgcagtctt cagagccaca ggaaccttga
     3241 gaatcattct aattatgatt tgctactcga aaaacatttt tcatcatcaa taataagaaa
     3301 ttaaaacggt atacatatag atgtttagta taacagttct tagtagttta aaagaacatt
     3361 tgtaacaaca ttccaacctt ttaataactt tagccttgct tacatttcta aaggagagcc
     3421 ttccagctgt ttttctgaac ccagtgatga atgtaaagtt cttaaaacca agttctaaga
     3481 atgttttaat gattacattt aacccaattt caagccaaat ttcctatacc acgagtaaaa
     3541 tatagttata attaggatag atctttggta aaataagata caattttaag atttgaattt
     3601 aaaagtagct aaattgtttt tttatgacag aacttttaag tttatagaat attaatacag
     3661 gacaaaactt gtaaatgtat ttaatttcct atgaatattt gataaaatat aaatgacatc
     3721 tatatgtaaa ttcatgaacc atgactctta tgaattatca gtgcacaaat tatgtttgat
     3781 gcacacttaa ggtcaaaagt ttagggtcag tttatttttt tttttatttt taattttatt
     3841 taataattta aataaaatta ttctgttcat caaggcacat ttattaaatg taacaataaa
     3901 ttgttaaata cttattaaat atttattttt tacttattgt atgcagtttc aattgaagtc
     3961 attattgctt ttattactat taccattaat aaaaataaca ttaataataa taataataat
     4021 aataataatt gatattacag tgatttctga aggatcacgt gactgaagac tggagtaatg
     4081 aagctgaaaa ctcatcattt aaatcactgg agtaaatgat tcaattaaat gatcatttaa
     4141 tgatcattct tttgaacagt tattttatag tgcaataaca tttcacagtt ttaccatttg
     4201 ttttgtattt ttgattcaat aaatgcagcc ttggtgagca ggagaagctt aatttaaaac
     4261 atttaaaact cctactgacc ccaaactttt gaccggtagt gtacatgtat tactgtatgt
     4321 tcaaactata tgcttttatt cataagtgtt tctattgctg tacatttgac tgtctgcatc
     4381 aggtgaagtg tatgaaagag tgtgtgttta ctccaccccg tgggaaagag cagtagctgc
     4441 aatgtgtgtg tgtgagtagt tagtgatgca tgagtgatcc actgaaccct gtccagcccc
     4501 ctgctgcagc tctcaacaca tcatgatagg tcacctctac tgggaggctt tcagcacagc
     4561 tctcacaact gatttatcag agagtactct gaaatggaga aattactttc atattaaaga
     4621 gcacacacac acacacacac acacacacac acacacacac acacacacac acacacacac
     4681 acacacctag ttcaccttat taacgtggtt aagcctttaa atgtcacttt aagctgtata
     4741 gaagtgtctt gaaaaatatc tattcaaata ttatttactg tcatcaagga aaagataaaa
     4801 taaatcagtt attagaaata agtttttaaa actattatac ttagaaatgt gctgaaacaa
     4861 tcttcctaga acagaacaga aattggggaa aaaataaaca aaaataaata aatctatcta
     4921 tctatagtgt gtataatgtg tgtagtgagt atgtttaaaa taggtcaact aatatgttca
     4981 taataaaaat acagcatcaa tggtttgggt gtttttagat ccaaaggtct taaacgtggt
     5041 tattgctggt tgttcagttt tttctaggct ctaagaaaaa tcattttaga aactgtaact
     5101 gtttccttat ccatcacctt tgggatgtag tggaacggga gatcaactgc atgatgctat
     5161 caagtcaata tggatcaaaa tctcaggaat atttccagga ccttattgaa tctatgccat
     5221 gaaggattaa ggcagttctg aaaggggaaa tgcggtttaa cccggtaata gcaaggtgta
     5281 cctaataaag tggccggtga gtgtatatat tgttattgtt tcaccttcag aaatgacata
     5341 tctagacaat agtttccttt ttctttttta attattgtac ttctgtaaga ttcttcaatg
     5401 ttcaatggtt ttcttatgtt aaaggcgcag agctgtagtt ttagtgaagt aacagcattc
     5461 cgttgaccaa agctgacaga tgaaggagag tcagttctga ccttgcagtt gtcagtcaca
     5521 attaagcctg gtttggttac aagttaaacg tagacctgct tgtggtcagg atcatccttt
     5581 aaaagaacta gatttgtttt taacaatttg taaccggtgt gctctgaagt gtcttaaata
     5641 aagcatatag gaagcaaagc agtgaaaaat gttccttata atctacgagc attagcttaa
     5701 atgtataaac aaaataatat taattgtgtg ttcagatcaa atataggtat tgttacacct
     5761 aaaataaacc cattgcttta aaaaataatt aaacacagtg tacaatatta gcatacttcc
     5821 tctgacatct ctaagtaaaa caagagcttt cacgatttac tcactgaccc atcttacagc
     5881 acttctctgt gaatttaggc catcacaaat gctgtgccca gttctctgac tgggtttgac
     5941 agtcgcccat ctgtgtccag cagtctgatg agctcatcct ctgtaaatct ggctggctgc
     6001 cgtgttgttt tgtgtctgtc cgtgagtctc tctggccttc atacactcct ctgctacttc
     6061 atagactgct gatgagtgca gaggccaaac cattagtgct tttctttttt tttagacctc
     6121 agtaatgcac aacaatgcaa aacaagtaga ttgagccttt tgttctttgg actgagcatt
     6181 tggttagaat agcttgaagg gatagttaga agatattttg aagggaaact aaaaacctgt
     6241 taccatttga catccaaaga aggaaaacta atatgattga gtggttacag atttccaaca
     6301 cttttcaaca tatattattt tgtgttccac taaagaaaca gtcaaatatt atactgtcaa
     6361 ccaaagtact gcacttgact caaaaaagct ctgcagcaag ttgcctctta ttctaggtcg
     6421 agtcaacctt tttttctctc tctctgtcgg ttcattcaag gttcagtaaa tgatgacaga
     6481 attttcagct tttattttgg gtgaactaac cctttatata tttgttgtaa gtttattttt
     6541 ctgtatatta taaatatatt agcaacataa ttctaagcta cacttgtacc caatcaaggg
     6601 tcacacttta ttttgatggt atgtttgttt gaatttaagt tacattgcat ctacatgtca
     6661 actaattctc attagattat aagaagactg ttaggttggg gttagtgtaa gttgacatgt
     6721 acttgcaaag tttctcagag tcagttaaat gtctgttttg tagcagtatt aacagatatt
     6781 aagcacactg tatattaata ctcaaatgaa ccgtcaaaat aaagtgttac cgtaaaaatt
     6841 atttccacat ctcaaggtag ctttatgagg aaagttttct ttaagattag tttcttcgaa
     6901 caaacttgca tgttaaatat gtgctcagat atggggcaca cagacaaaat gtttattgga
     6961 cgaataaccc ctttaacagt taaccagcat tttttaaagg gatagtcaga agatattttg
     7021 aaggaaacta aaaacctgta accattcgac aatcgaagaa ggtaaaagaa atatgataga
     7081 gtagttacag tttttcaaca ctctttataa tatattgttt agtattcaac tgaagaaata
     7141 gtcaaacaca ctgtaaaaaa agttggttta aattgactca actttcaact caagtatagt
     7201 acttgacttg atttaagttg agttaactag actaaactaa aagcactaaa agcactgcgg
     7261 caagtcgcct ttcaatttta ggttgagtaa attattatta ttttttttta cagttcaggt
     7321 tcagtaaatg ataacataat tttcagtttt gggtgaacta tccctttata tatttgttgt
     7381 aagttaattt ttctgtaaat tataaattaa ttagcaaaaa agctgcactt cttcaccatt
     7441 aattaattaa tacatctcaa agtagcttta taaggaaagt tttctttaat tatgtttctt
     7501 tgcatgttaa aattgtgctc agatatggag cacacaagca aaatgtttat tggatgaata
     7561 acccctttaa catttaacca gcatgagttc aagtaaacaa atcaattcct atgatttttc
     7621 tgctaaaccg atataaatca gtgctgctga agagcatgtg tgtgatgttc tcagatcagc
     7681 acaaaagagg gagcgccagt ttcttttaat aatgaagaat tacgtaatca cctcctgtgt
     7741 tccctgaaac cattaaactc ccagacagaa caaaactgtg actgagaaag cagagcaact
     7801 ccggagtaac tcagaagatc tgcaaaccaa ctctttagca tgaggtaaga gagaattaaa
     7861 gtccaatatc agtgctgatg ccatctcagt ctctctattt ctgtgttcca gcagggtatg
     7921 aataggattt tctctggctc actgtgaaaa gaggcagtca ggagactggc tggacaagca
     7981 caaacaattg cgggctcaca aacaccccct cggcctttgc cctccccatc agaccccccc
     8041 tctctctttc accccccaca tcaactctat aaaaacccag agttcacccg cactcgatct
     8101 cattctcctc caccatggag tcccagcgtt cctcgtcgac ggtaccgcgg gcccgggatc
     8161 cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     8221 ggttaattca agtttgtaca aaaaagttga acgagaaacg taaaatgata taaatatcaa
     8281 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
     8341 gcagtcacta tgaaccaact acttagatgg tattagtgac ctgtagaatt cgagctctag
     8401 agctgcaggg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     8461 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     8521 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     8581 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     8641 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     8701 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     8761 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     8821 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     8881 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     8941 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     9001 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     9061 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     9121 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     9181 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     9241 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     9301 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     9361 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     9421 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     9481 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     9541 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     9601 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     9661 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     9721 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     9781 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     9841 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     9901 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     9961 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    10021 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    10081 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    10141 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
