LOCUS       sequence-202447-        7136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP from GFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7136)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 7136)
  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"
COMMENT     Sequence Label: pAAV.GFAP.eGFP.WPRE.hGH
FEATURES             Location/Qualifiers
     source          1..7136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     misc_feature    190..307
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      2564..2573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2570..2572,2573..2575,2576..3289)
                     /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(2570..2572,2573..2575,2576..3286)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2615..2636)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2876..2895)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3223..3244
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3307..3895
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3360..3380)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3778..3789)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3927..4403
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4126..4145)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4443..4583
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4443..4572
                     /label=AAV2 ITR
     rep_origin      4658..5113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4745..4764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4955..4976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5130..5149)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5249..5271
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5309..5327)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5395..5499
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5500..5568,5569..6360)
                     /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      6531..7119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7020..7039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
      181 gcacgcgttt tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
      241 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
      301 aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta acccacctcc
      361 ctctctgtgc tgggactcac agagggagac ctcaggaggc agtctgtcca tcacatgtcc
      421 aaatgcagag cataccctgg gctgggcgca gtggcgcaca actgtaattc cagcactttg
      481 ggaggctgat gtggaaggat cacttgagcc cagaagttct agaccagcct gggcaacatg
      541 gcaagaccct atctctacaa aaaaagttaa aaaatcagcc acgtgtggtg acacacacct
      601 gtagtcccag ctattcagga ggctgaggtg aggggatcac ttaaggctgg gaggttgagg
      661 ctgcagtgag tcgtggttgc gccactgcac tccagcctgg gcaacagtga gaccctgtct
      721 caaaagacaa aaaaaaaaaa aaaaaaaaaa agaacatatc ctggtgtgga gtaggggacg
      781 ctgctctgac agaggctcgg gggcctgagc tggctctgtg agctggggag gaggcagaca
      841 gccaggcctt gtctgcaagc agacctggca gcattgggct ggccgccccc cagggcctcc
      901 tcttcatgcc cagtgaatga ctcaccttgg cacagacaca atgttcgggg tgggcacagt
      961 gcctgcttcc cgccgcaccc cagcccccct caaatgcctt ccgagaagcc cattgagcag
     1021 ggggcttgca ttgcacccca gcctgacagc ctggcatctt gggataaaag cagcacagcc
     1081 ccctaggggc tgcccttgct gtgtggcgcc accggcggtg gagaacaagg ctctattcag
     1141 cctgtgccca ggaaagggga tcaggggatg cccaggcatg gacagtgggt ggcagggggg
     1201 gagaggaggg ctgtctgctt cccagaagtc caaggacaca aatgggtgag gggactgggc
     1261 agggttctga ccctgtggga ccagagtgga gggcgtagat ggacctgaag tctccaggga
     1321 caacagggcc caggtctcag gctcctagtt gggcccagtg gctccagcgt ttccaaaccc
     1381 atccatcccc agaggttctt cccatctctc caggctgatg tgtgggaact cgaggaaata
     1441 aatctccagt gggagacgga ggggtggcca gggaaacggg gcgctgcagg aataaagacg
     1501 agccagcaca gccagctcat gtgtaacggc tttgtggagc tgtcaaggcc tggtctctgg
     1561 gagagaggca cagggaggcc agacaaggaa ggggtgacct ggagggacag atccaggggc
     1621 taaagtcctg ataaggcaag agagtgccgg ccccctcttg ccctatcagg acctccactg
     1681 ccacatagag gccatgattg acccttagac aaagggctgg tgtccaatcc cagcccccag
     1741 ccccagaact ccagggaatg aatgggcaga gagcaggaat gtgggacatc tgtgttcaag
     1801 ggaaggactc caggagtctg ctgggaatga ggcctagtag gaaatgaggt ggcccttgag
     1861 ggtacagaac aggttcattc ttcgccaaat tcccagcacc ttgcaggcac ttacagctga
     1921 gtgagataat gcctgggtta tgaaatcaaa aagttggaaa gcaggtcaga ggtcatctgg
     1981 tacagccctt ccttcccttt tttttttttt ttttttgtga gacaaggtct ctctctgttg
     2041 cccaggctgg agtggcgcaa acacagctca ctgcagcctc aacctactgg gctcaagcaa
     2101 tcctccagcc tcagcctccc aaagtgctgg gattacaagc atgagccacc ccactcagcc
     2161 ctttccttcc tttttaattg atgcataata attgtaagta ttcatcatgg tccaaccaac
     2221 cctttcttga cccaccttcc tagagagagg gtcctcttgc ttcagcggtc agggccccag
     2281 acccatggtc tggctccagg taccacctgc ctcatgcagg agttggcgtg cccaggaagc
     2341 tctgcctctg ggcacagtga cctcagtggg gtgaggggag ctctccccat agctgggctg
     2401 cggcccaacc ccaccccctc aggctatgcc agggggtgtt gccaggggca cccgggcatc
     2461 gccagtctag cccactcctt cataaagccc tcgcatccca ggagcgagca gagccagagc
     2521 aggttggaga ggagacgcat cacctccgct gctcgccggg atggccgcca tggtgagcaa
     2581 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2641 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2701 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2761 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2821 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2881 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2941 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3001 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3061 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3121 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     3181 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3241 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat aagctagctt
     3301 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3361 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3421 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3481 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3541 cccactggtt ggggcattgc caccacctgt cagctccttt ccggaacttt cgctttcccc
     3601 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3661 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     3721 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3781 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3841 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     3901 taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc
     3961 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     4021 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     4081 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     4141 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     4201 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     4261 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     4321 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     4381 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc
     4441 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4501 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     4561 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     4621 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     4681 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     4741 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4801 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4861 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4921 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4981 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     5041 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5101 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5161 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5221 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5281 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5341 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5401 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5461 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5521 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5581 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5641 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5701 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5761 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5821 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5881 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5941 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6001 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6061 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6121 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6181 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6241 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6301 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6361 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6421 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6481 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6541 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6601 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6661 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6721 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6781 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6841 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6901 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6961 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7021 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7081 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggc
//
