LOCUS       sequence-450679-       10185 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 10185)
  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 10185)
  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..10185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(919..946)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1038..1124)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1173..1195
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1187..1204
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1188..1204
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1244..1339
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1341..1359
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8810..8826)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8810..8826)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(8863..8881)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    8882..9005
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(9078..9096)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9101..9117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9101..9117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9219..9238)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             9230..10039
                     /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(9297..9316)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
      901 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
      961 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     1021 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     1081 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     1141 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc
     1201 cagtcttaag ctcgggcccg agttaacgct accatggagc tccaaataat gattttattt
     1261 tgactgatag tgacctgttc gttgcaacaa attgataagc aatgcttttt tataatgcca
     1321 actttgtata gaaaagttgg taatacgact cactataggg cgaattgggt accgggcccc
     1381 ccctcgaggt aaggactgag gtgatgcaca gtgttgggta tgttactttc aataagttat
     1441 tctttgctaa ttattagtca tttgaattgt attgaaatta catttttagt tactttgcct
     1501 aaaaaaaaac ttgataagta atcgaattac atacttgaag tccacataaa tctcattcat
     1561 tcattccttc tcttttcgac ttagtccctt tattaatcag gggtttctac agcggaacga
     1621 acctcatatg tttcaattta gtgactattt tttataaatt attgacaatg tttttgttgc
     1681 ttgttcaaac tacttattta aaatgagctg aaactgcaca attattgata tttcattgga
     1741 acagcttaat tttttttatg tttaatccac ttaaataaaa ttccaattat ataatattcc
     1801 aattataaag ttacttaatc aatttgtatt aggacaatat gaatgaattg tgtggaaccc
     1861 tgcatttttt acagtgaacc tttccttgtt tcttaaacat gggtgatttt cggagaaata
     1921 aaatttgaca atatctgaaa tgtaaaaaga aaatattcag ttactctgcc tttctaaatg
     1981 catagtttat attatcatca gagttgtaaa gtaactagtt acaaatactc aaattactgt
     2041 aattgagtag tttttttcag gaattgtaat ttactaagta gctaaaaaat gtgtactttt
     2101 actttccctt gagtacatta ttaatgctgt aacggtactt ttactccact actttccttc
     2161 agcctgcact cactactttg ttttatcttg tctatgagga ttagaaaaat caatcctgtg
     2221 attccaatca aatcgcacat ccaaagtctt tacaaaagca ttttgtttgg atgtgaaaat
     2281 cgggttgatg tcagaactca atgtcagccc aacgtcaatg tccaacctaa aatcaaccta
     2341 atatcaacgt ctaatgatgt tacagcttga cgttgtgtgg gcgttaccat tatttgtcag
     2401 acattggatt ttagttgcca tacctgataa ataaatgtca gtatttgatg tccgtatgat
     2461 cttggattta gatgttggct cgacattgga ttttggtcac ctgacataac aacctaaatc
     2521 tatcctaata ataacgtctt atgacgttgt gtgccaacca tttttgttta ttaaaaattt
     2581 atataaacat ttacaataat atattatcca tgtgatttaa tagaaatgtt atttgttttt
     2641 tttctccccc atatttctga atgacttaaa gcttttatta tcaggataag gttgtggttt
     2701 ttattttttg cttgctaact tcaattgcta ttgcatttac atttatgcaa atggggagac
     2761 gcggtgggtc agttggcgtt tctgtgtgga gtttgcatgt tctccccgtg ttcgcgtggg
     2821 ttttctctgg gtgctctggt ttcccctaaa gacatgtagt ataggtgaat tgggtaagct
     2881 aaattgtccg tagtgtgtat gtgtgaatgt atggatgttt tcctgtgatg ggttgcagct
     2941 gaaagggcat ccgctgtgta aaacatatgc tggataagtt ggcaattcat ttcgctgtgg
     3001 taaccccaga ttaatatcgg aactacgcca aaaagaaaat gaatgaatga gtgaattaat
     3061 gcaaatggct catccttcca cctaattcca gtgccttgca ttgcattcaa gctctacaat
     3121 taatcagttc ctttattttc actttccaaa ctgaatccaa caactcatga ggaccccagg
     3181 gtgccttgat ttaccagtgt catgatgaac acaatctacc ccactttaac ccactcgtca
     3241 acataaatac tcccttaaag ggtgtacttg tgtctgagta gaagtgtctg ctgggttgga
     3301 ggtgtcgcac attccaggac acgtcaagaa gaacgctaag agcaacagga gaacaatgtc
     3361 acaaaaaagg tcaaagaaac gccatccatt agccagcttt ataaacagct gtctcgtggc
     3421 taaagaagtt ctcaagaata accaaccttg catttttcca cattagtttt acaccgcagc
     3481 aaagtcacca tgtggattta tagcatccaa accccatttt caatgagagg cttcacattt
     3541 ctcgctctca ttcaatccca cactgcctct cttttcagac taaatcctga taggatgagt
     3601 ggtatggggg cggggagggg gcacaatacc tcagtccagc ttccacaaag actcaacaca
     3661 cccatgcaga tgtcctgttg ttggggaagt gcatttgtaa ccaaatttca aactttcaac
     3721 tttcatttga agcagtggag actgctccat tcctattcag gatcaattta ttgaaatatg
     3781 acaacattta gaataacttt gttatatgtt attatgtttt ataatgcaat tttgtttctg
     3841 tgacatacat ttttttattt agtgcagtct tcagagccac aggaaccttg agaatcattc
     3901 taattatgat ttgctactcg aaaaacattt ttcatcatca ataataagaa attaaaacgg
     3961 tatacatata gatgtttagt ataacagttc ttagtagttt aaaagaacat ttgtaacaac
     4021 attccaacct tttaataact ttagccttgc ttacatttct aaaggagagc cttccagctg
     4081 tttttctgaa cccagtgatg aatgtaaagt tcttaaaacc aagttctaag aatgttttaa
     4141 tgattacatt taacccaatt tcaagccaaa tttcctatac cacgagtaaa atatagttat
     4201 aattaggata gatctttggt aaaataagat acaattttaa gatttgaatt taaaagtagc
     4261 taaattgttt ttttatgaca gaacttttaa gtttatagaa tattaataca ggacaaaact
     4321 tgtaaatgta tttaatttcc tatgaatatt tgataaaata taaatgacat ctatatgtaa
     4381 attcatgaac catgactctt atgaattatc agtgcacaaa ttatgtttga tgcacactta
     4441 aggtcaaaag tttagggtca gtttattttt tttttatttt taattttatt taataattta
     4501 aataaaatta ttctgttcat caaggcacat ttattaaatg taacaataaa ttgttaaata
     4561 cttattaaat atttattttt tacttattgt atgcagtttc aattgaagtc attattgctt
     4621 ttattactat taccattaat aaaaataaca ttaataataa taataataat aataataatt
     4681 gatattacag tgatttctga aggatcacgt gactgaagac tggagtaatg aagctgaaaa
     4741 ctcatcattt aaatcactgg agtaaatgat tcaattaaat gatcatttaa tgatcattct
     4801 tttgaacagt tattttatag tgcaataaca tttcacagtt ttaccatttg ttttgtattt
     4861 ttgattcaat aaatgcagcc ttggtgagca ggagaagctt aatttaaaac atttaaaact
     4921 cctactgacc ccaaactttt gaccggtagt gtacatgtat tactgtatgt tcaaactata
     4981 tgcttttatt cataagtgtt tctattgctg tacatttgac tgtctgcatc aggtgaagtg
     5041 tatgaaagag tgtgtgttta ctccaccccg tgggaaagag cagtagctgc aatgtgtgtg
     5101 tgtgagtagt tagtgatgca tgagtgatcc actgaaccct gtccagcccc ctgctgcagc
     5161 tctcaacaca tcatgatagg tcacctctac tgggaggctt tcagcacagc tctcacaact
     5221 gatttatcag agagtactct gaaatggaga aattactttc atattaaaga gccacacaca
     5281 cacacacaca cacacacaca cacacacaca cacacacaca cacacacaca cacctagttc
     5341 accttattaa cgtggttaag cctttaaatg tcactttaag ctgtatagaa gtgtcttgaa
     5401 aaatatctat tcaaatatta tttactgtca tcaaggaaaa gataaaataa atcagttatt
     5461 agaaataagt ttttaaaact attatactta gaaatgtgct gaaacaatct tcctagaaca
     5521 gaacagaaat tggggaaaaa ataaacaaaa ataaataaat ctatctatct atagtgtgta
     5581 taatgtgtgt agtgagtatg tttaaaatag gtcaactaat atgttcataa taaaaataca
     5641 gcatcaatgg tttgggtgtt tttagatcca aaggtcttaa acgtggttat tgctggttgt
     5701 tcagtttttt ctaggctcta agaaaaatca ttttagaaac tgtaactgtt tccttatcca
     5761 tctgaattcc acctttggga tgtagtggaa cgggagatca actgcatgat gctatcaagt
     5821 caatatggat caaaatctca ggaatatttc caggacctta ttgaatctat gccatgaagg
     5881 attaaggcag ttctgaaagg ggaaatgcgg tttaacccgg taatagcaag gtgtacctaa
     5941 taaagtggcc ggtgagtgta tatattgtta ttgtttcacc ttcagaaatg acatatctag
     6001 acaatagttt cctttttctt ttttaattat tgtacttctg taagattctt caatgttcaa
     6061 tggttttctt atgttaaagg cgcagagctg tagttttagt gaagtaacag cattccgttg
     6121 accaaagctg acagatgaag gagagtcagt tctgaccatg cagttgtcag tcacaattaa
     6181 gcctggtttg gttacaagtt aaacgtagac ctgcttgtgg tcaggatcat cctttaaaag
     6241 aactagattt gtttttaaca atttgtaacc ggtgtgctct gaagtgtctt aaataaagca
     6301 tataggaagc aaagcagtga aaaatgttcc ttataatcta cgagcattag cttaaatgta
     6361 taaacaaaat aatattaatt gtgtgttcag atcaaatata ggtattgtta cacctaaaat
     6421 aaacccattg ctttaaaaaa taattaaaca cagtgtacaa tattagcata cttcctctga
     6481 catctctaag taaaacaaga gctttcacga tttactcact gacccatctt acagcacttc
     6541 tctgtgaatt taggccatca caaatgctgt gcccagttct ctgactgggt ttgacagtcg
     6601 cccatctgtg tccagcagtc tgatgagctc atcctctgta aatctggctg gctgccgtgt
     6661 tgttttgtgt ctgtccgtga gtctctctgg ccttcataca ctcctctgct acttcataga
     6721 ctgctgatga gtgcagaggc caaaccatta gtgctttcct ttttttagac ctcagtaatg
     6781 cacaacaatg caaaacaagt agattgagcc ttttgttctt tggactgagc atttggttag
     6841 aatagcttga agggatagtt agaagatatt ttgaagggaa actaaaaacc tgttaccatt
     6901 tgacatccaa agaaggaaaa ctaatatgat tgagtggtta cagatttcca acacttttca
     6961 acatatatta ttttgtgttc cactaaagaa acagtcagat attatactgt caaccaaagt
     7021 actgcacttg actcaaaaaa gctctgcagc aagttgcctc ttattctagg tcgagtcaac
     7081 ctttttttct ctctctctgt cggttcattc aaggttcagt aaatgatgac agaattttca
     7141 gcttttattt tgggtgaact aaccctttat atatttgttg taagtttatt tttctgtata
     7201 ttataaatat attagcaaca taattctaag ctacacttgt acccaatcaa gggtcacact
     7261 ttattttgat ggtatgtttg tttgaattta agttacattg catctacatg tcaactaatt
     7321 ctcattagat tataagaaga ctgttaggtt ggggctagtg taagttgaca tgtacttgca
     7381 aagtttctca gagtcagtta aatgtctgtt ttgtagcagt attaacagat attaagcaca
     7441 ctgtatatta atactcaaat gaaccgtcaa aataaagtgt taccgtaaaa attatttcca
     7501 catctcaagg tagctttatg aggaaagttt tctttaagat tagtttcttc gaacaaactt
     7561 gcatgttaaa tatgtgctca gatatggggc acacagacaa aatgtttatt ggacgaataa
     7621 cccctttaac agttaaccag cattttttaa agggatagtc agaagatatt ttgaaggaaa
     7681 ctaaaaacct gtaaccattc gacaatcgaa gaaggtaaaa gaaatatgat agagtagtta
     7741 cagtttttca acactcttta taatatattg tttagtattc aactgaagaa atagtcaaac
     7801 acactgtaaa aaaagttggt ttaaattgac tcaactttca actcaagtat agtacttgac
     7861 ttgatttaag ttgagttaac tagactaaac taaaagcact aaaagcactg cggcaagtcg
     7921 cctttcaatt ttaggttgag taaattatta ttattttttt ttacagttca ggttcagtaa
     7981 atgataacat aattttcagt tttgggtgaa ctatcccttt atatatttgt tgtaagttaa
     8041 tttttctgta aattataaat taattagcaa aaaagctgca cttcttcacc attaattaat
     8101 taatacatct caaagtagct ttataaggaa agttttcttt aattatgttt ctttgcatgt
     8161 taaaattgtg ctcagatatg gagcacacaa gcaaaatgtt tattggatga ataacccctt
     8221 taacatttaa ccagcatgag ttcaagtaaa caaatcaatt cctatgattt ttctgctaaa
     8281 ccgatataaa tcagtgctgc tgaagagcat gtgtgtgatg ttctcagatc agcacaaaag
     8341 agggagcgcc agtttctttt aataatgaag aattacgtaa tcacctcctg tgttccctga
     8401 aaccattaaa ctcccagaca gaacaaaact gtgactgaga aagcagagca actccggagt
     8461 aactcagaag atctgcaaac caactcttta gcatgaggta agagagaatt aaagtccaat
     8521 atcagtgctg atgccatctc agtctctcta tttctgtgtt ccagcagggt atgaatagga
     8581 ttttctctgg ctcactgtga aaagaggcag tcaggagact ggctggacaa gcacaaacaa
     8641 ttgcgggctc acaaacaccc cctcggcctt tgccctcccc atcagacccc ccctctctct
     8701 ttcacccccc acatcaactc tataaaaacc cagagttcac ccgcactcga tctcattctc
     8761 ctccaccatg gagtcccagc gttcctcgtc gacggtaccg cgggcccggg atccactagt
     8821 tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     8881 tcaagtttgt acaaaaaagt tgaacgagaa acgtaaaatg atataaatat caatatatta
     8941 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatgcagtca
     9001 ctatgaacca actacttaga tggtattagt gacctgtaga attcgagctc tagagctgca
     9061 gggcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     9121 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     9181 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     9241 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     9301 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     9361 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     9421 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     9481 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     9541 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     9601 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     9661 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     9721 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     9781 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     9841 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     9901 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     9961 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
    10021 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
    10081 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
    10141 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
