LOCUS       sequence-201071-        7147 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 7147)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 7147)
  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..7147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     misc_feature    146..263
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      2520..2529
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2526..2528,2529..2531,2532..3245)
                     /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(2526..2528,2529..2531,2532..3242)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2571..2592)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2832..2851)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3179..3200
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3263..3851
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3316..3336)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3734..3745)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3883..4359
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4082..4101)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4399..4539
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4399..4528
                     /label=AAV2 ITR
     rep_origin      4614..5069
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4701..4720)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4911..4932
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5086..5105)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5205..5227
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5265..5283)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5351..5455
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5456..5524,5525..6316)
                     /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      6487..7075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6976..6995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7137..7147,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttttta aaagaaaagg ggggattggg gggtacagtg
      181 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
      241 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt
      301 taattaaccc acctccctct ctgtgctggg actcacagag ggagacctca ggaggcagtc
      361 tgtccatcac atgtccaaat gcagagcata ccctgggctg ggcgcagtgg cgcacaactg
      421 taattccagc actttgggag gctgatgtgg aaggatcact tgagcccaga agttctagac
      481 cagcctgggc aacatggcaa gaccctatct ctacaaaaaa agttaaaaaa tcagccacgt
      541 gtggtgacac acacctgtag tcccagctat tcaggaggct gaggtgaggg gatcacttaa
      601 ggctgggagg ttgaggctgc agtgagtcgt ggttgcgcca ctgcactcca gcctgggcaa
      661 cagtgagacc ctgtctcaaa agacaaaaaa aaaaaaaaaa aaaaaaagaa catatcctgg
      721 tgtggagtag gggacgctgc tctgacagag gctcgggggc ctgagctggc tctgtgagct
      781 ggggaggagg cagacagcca ggccttgtct gcaagcagac ctggcagcat tgggctggcc
      841 gccccccagg gcctcctctt catgcccagt gaatgactca ccttggcaca gacacaatgt
      901 tcggggtggg cacagtgcct gcttcccgcc gcaccccagc ccccctcaaa tgccttccga
      961 gaagcccatt gagcaggggg cttgcattgc accccagcct gacagcctgg catcttggga
     1021 taaaagcagc acagccccct aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga
     1081 acaaggctct attcagcctg tgcccaggaa aggggatcag gggatgccca ggcatggaca
     1141 gtgggtggca gggggggaga ggagggctgt ctgcttccca gaagtccaag gacacaaatg
     1201 ggtgagggga ctgggcaggg ttctgaccct gtgggaccag agtggagggc gtagatggac
     1261 ctgaagtctc cagggacaac agggcccagg tctcaggctc ctagttgggc ccagtggctc
     1321 cagcgtttcc aaacccatcc atccccagag gttcttccca tctctccagg ctgatgtgtg
     1381 ggaactcgag gaaataaatc tccagtggga gacggagggg tggccaggga aacggggcgc
     1441 tgcaggaata aagacgagcc agcacagcca gctcatgtgt aacggctttg tggagctgtc
     1501 aaggcctggt ctctgggaga gaggcacagg gaggccagac aaggaagggg tgacctggag
     1561 ggacagatcc aggggctaaa gtcctgataa ggcaagagag tgccggcccc ctcttgccct
     1621 atcaggacct ccactgccac atagaggcca tgattgaccc ttagacaaag ggctggtgtc
     1681 caatcccagc ccccagcccc agaactccag ggaatgaatg ggcagagagc aggaatgtgg
     1741 gacatctgtg ttcaagggaa ggactccagg agtctgctgg gaatgaggcc tagtaggaaa
     1801 tgaggtggcc cttgagggta cagaacaggt tcattcttcg ccaaattccc agcaccttgc
     1861 aggcacttac agctgagtga gataatgcct gggttatgaa atcaaaaagt tggaaagcag
     1921 gtcagaggtc atctggtaca gcccttcctt cccttttttt tttttttttt ttgtgagaca
     1981 aggtctctct ctgttgccca ggctggagtg gcgcaaacac agctcactgc agcctcaacc
     2041 tactgggctc aagcaatcct ccagcctcag cctcccaaag tgctgggatt acaagcatga
     2101 gccaccccac tcagcccttt ccttcctttt taattgatgc ataataattg taagtattca
     2161 tcatggtcca accaaccctt tcttgaccca ccttcctaga gagagggtcc tcttgcttca
     2221 gcggtcaggg ccccagaccc atggtctggc tccaggtacc acctgcctca tgcaggagtt
     2281 ggcgtgccca ggaagctctg cctctgggca cagtgacctc agtggggtga ggggagctct
     2341 ccccatagct gggctgcggc ccaaccccac cccctcaggc tatgccaggg ggtgttgcca
     2401 ggggcacccg ggcatcgcca gtctagccca ctccttcata aagccctcgc atcccaggag
     2461 cgagcagagc cagagcaggt tggagaggag acgcatcacc tccgctgctc gccgggatgg
     2521 ccgccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2581 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2641 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2701 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2761 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2821 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2881 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2941 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3001 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3061 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3121 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3181 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3241 agtaataagc tagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3301 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3361 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3421 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3481 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3541 aactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3601 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3661 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     3721 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3781 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3841 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3901 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3961 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     4021 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     4081 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     4141 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     4201 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     4261 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     4321 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4381 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4441 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4501 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4561 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4621 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4681 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4741 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4801 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4861 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4921 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     4981 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5041 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5101 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5161 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5221 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5281 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5341 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5401 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5461 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5521 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5581 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5641 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5701 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5761 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5821 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5881 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5941 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6001 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6061 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6121 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6181 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6241 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6301 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6361 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6421 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6481 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6541 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6601 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6661 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6721 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6781 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6841 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6901 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6961 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7021 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7081 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg
     7141 caggcag
//
