LOCUS       sequence-189835-        5673 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 5673)
  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 5673)
  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..5673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          1000..1132
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1115..1134
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1144..1153
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1150..1152,1153..1155,1156..1869)
                     /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(1150..1152,1153..1155,1156..1866)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1195..1216)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1456..1475)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1803..1824
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1936..1956)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2354..2365)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2432..2449)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2438..2645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2733..2862
                     /label=AAV2 ITR
     primer_bind     complement(2881..2898)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2881..2897)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2890..2912)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3039..3494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3126..3145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3336..3357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3521..3625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3626..3694,3695..4486)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4657..5245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5146..5165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5533..5554
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5569..5574,5575..5592,5593..5599)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5607..5623
                     /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     5612..5634
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5631..5647
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5631..5647
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg gaattcgccc ttaagctagc cgagctcggt accagatcta acatatcctg
      241 gtgtggagta ggggacgctg ctctgacaga ggctcggggg cctgagctgg ctctgtgagc
      301 tggggaggag gcagacagcc aggccttgtc tgcaagcaga cctggcagca ttgggctggc
      361 cgccccccag ggcctcctct tcatgcccag tgaatgactc accttggcac agacacaatg
      421 ttcggggtgg gcacagtgcc tgcttcccgc cgcaccccag cccccctcaa atgccttccg
      481 agaagcccat tgagcagggg gcttgcattg caccccagcc tgacagcctg gcatcttggg
      541 ataaaagcag cacagccccc taggggctgc ccttgctgtg tggcgccacc ggcggtggag
      601 aacaaggctc tattcagcct gtgcccagga aaggggatca ggggatgccc aggcatggac
      661 agtgggtggc agggggggag aggagggctg tctgcttccc agaagtccaa ggacacaaat
      721 gggtgagggg agagctctcc ccatagctgg gctgcggccc aaccccaccc cctcaggcta
      781 tgccaggggg tgttgccagg ggcacccggg catcgccagt ctagcccact ccttcataaa
      841 gccctcgcat cccaggagcg agcagagcca gagcaggttg gagaggagac gcatcacctc
      901 cgctgctcgc ggggatcctc tagagtcgac ggatccgggg aattccccag tttcaggatc
      961 caccatggct gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga
     1021 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     1081 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccag
     1141 gcggccgcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1201 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1261 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1321 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1381 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1441 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1501 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1561 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1621 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1681 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1741 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1801 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1861 tacaagtaat aagcttggat ccaatcaacc tctggattac aaaatttgtg aaagattgac
     1921 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1981 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2041 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2101 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2161 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2221 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2281 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2341 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2401 tctgcggcct cttccgcgtc ttcgagatct gcctcgactg tgccttctag ttgccagcca
     2461 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2521 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2581 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2641 ggggactcga gttaagggcg aattcccgat aaggatcttc ctagagcatg gctacgtaga
     2701 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     2761 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     2821 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt aacctaattc
     2881 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     2941 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     3001 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     3061 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3121 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3181 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3241 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccccgat agacggtttt
     3301 tcgccctttg acgctggagt tcacgttcct caatagtgga ctcttgttcc aaactggaac
     3361 aacactcaac cctatctcgg tctattcttt tgatttataa gggatttttc cgatttcggc
     3421 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3481 aacgtttata atttcaggtg gcatctttcg gggaaatgtg cgcggaaccc ctatttgttt
     3541 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3601 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3661 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3721 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaatagtgg
     3781 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3841 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3901 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3961 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4021 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4081 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4141 aaacgacgag cgtgacacca cgatgcctgt agtaatggta acaacgttgc gcaaactatt
     4201 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4261 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4321 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4381 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4441 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4501 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4561 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4621 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4681 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4741 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4801 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4861 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4921 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4981 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5041 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5101 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5161 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5221 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5281 cttttgctgc ggttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5341 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5401 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     5461 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5521 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     5581 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     5641 ctatgaccat gattacgcca gatttaatta agg
//
