LOCUS       sequence-199087-        5683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5683)
  AUTHORS   Ortinski PI, Dong J, Mungenast A, Yue C, Takano H, Watson DJ, Haydon
            PG, Coulter DA
  TITLE     Selective induction of astrocytic gliosis generates deficits in 
            neuronal inhibition.
  JOURNAL   Nat Neurosci. 2010 May;13(5):584-91. doi: 10.1038/nn.2535. Epub 2010
            Apr 25.
  PUBMED    20418874
REFERENCE   2  (bases 1 to 5683)
  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"; doi: "10.1038/nn.2535"; 
            journalName: "Nat Neurosci"; date: "2010-05"; volume: "13"; issue: 
            "5"; pages: "584-91"
COMMENT     Sequence Label: pAAV.GFA104.PI.eGFP.WPRE.bGH
FEATURES             Location/Qualifiers
     source          1..5683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          816..948
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     931..950
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      960..969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(966..968,969..971,972..1685)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(966..968,969..971,972..1682)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1011..1032)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1272..1291)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1619..1640
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1752..1772)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2170..2181)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2248..2265)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2254..2461
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2549..2678
                     /label=AAV2 ITR
     primer_bind     complement(2697..2714)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2697..2713)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2706..2728)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2855..3310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2942..2961)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3152..3173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3336..3440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3441..3509,3510..4301)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4472..5060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4961..4980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5348..5369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5384..5389,5390..5407,5408..5414)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5422..5438
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5427..5449
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5446..5462
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5446..5462
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5500..5629
                     /label=AAV2 ITR
ORIGIN
        1 agatcggaat tcgcccttaa gctagccgag ctcggtacca gatctaacat atcctggtgt
       61 ggagtagggg acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg
      121 gaggaggcag acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc
      181 ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg
      241 gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa
      301 gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa
      361 aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca
      421 aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg
      481 ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt
      541 gaggggagag ctctccccat agctgggctg cggcccaacc ccaccccctc aggctatgcc
      601 agggggtgtt gccaggggca cccgggcatc gccagtctag cccactcctt cataaagccc
      661 tcgcatccca ggagcgagca gagccagagc aggttggaga ggagacgcat cacctccgct
      721 gctcgcgggg atcctctaga gtcgacggat ccggggaatt ccccagtttc aggatccacc
      781 atggctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg
      841 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
      901 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccaggcgg
      961 ccgccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1021 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1081 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1141 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1201 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1261 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1321 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1381 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1441 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1501 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1561 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1621 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1681 agtaataagc ttggatccaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1741 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1801 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     1861 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     1921 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     1981 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2041 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2101 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2161 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2221 cggcctcttc cgcgtcttcg agatctgcct cgactgtgcc ttctagttgc cagccatctg
     2281 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2341 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2401 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2461 actcgagtta agggcgaatt cccgataagg atcttcctag agcatggcta cgtagataag
     2521 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     2581 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2641 tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc taattcactg
     2701 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2761 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2821 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     2881 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2941 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3001 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3061 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3121 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3181 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3241 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3301 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3361 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3421 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3481 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3541 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3601 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3661 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3721 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3781 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3841 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3901 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3961 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4021 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4081 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4141 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4201 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4261 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4321 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4381 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5101 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5161 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5221 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5281 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5341 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     5401 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     5461 accatgatta cgccagattt aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact
     5521 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     5581 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg
     5641 attaacccgc catgctactt atctacgtag ccatgctcta gga
//
