LOCUS       sequence-379942-        4871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector plasmid expressing enhanced green fluorescent protein 
            (eGFP) under the mouse phosphoglycerate kinase (PGK) promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4871)
  AUTHORS   Nieuwenhuis B, Laperrousaz E, Tribble JR, Verhaagen J, Fawcett JW, 
            Martin KR, Williams PA, Osborne A
  TITLE     Improving adeno-associated viral (AAV) vector-mediated transgene 
            expression in retinal ganglion cells: comparison of five promoters.
  JOURNAL   Gene Ther. 2023 Jan 13. doi: 10.1038/s41434-022-00380-z.
  PUBMED    36635457
REFERENCE   2  (bases 1 to 4871)
  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: "Gene Ther. 
            2023 Jan 13. doi: 10.1038/s41434-022-00380-z."
COMMENT     Sequence Label: AAV-PGK-eGFP
FEATURES             Location/Qualifiers
     source          1..4871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   9..138
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        188..687
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(223..246)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     620..639
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      843..852
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(849..851,852..854,855..1568)
                     /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(849..851,852..854,855..1565)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(894..915)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1155..1174)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1502..1523
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1626..2102
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1825..1844)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2142..2282
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2142..2271
                     /label=AAV2 ITR
     rep_origin      2357..2812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2444..2463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2654..2675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2829..2848)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2948..2970
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3008..3026)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3094..3198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3199..3267,3268..4059)
                     /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"
     primer_bind     complement(3417..3436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4230..4818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4719..4738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg
       61 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
      121 ccatcactag gggttcctgc ggccgcacgc gcgatcacga gactagcctc gacgatggtc
      181 gagtaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
      241 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
      301 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
      361 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
      421 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
      481 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
      541 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
      601 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
      661 tcctcttcct catctccggg cctttcgacc tctagcggga tcggattcga atcccggccg
      721 ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag
      781 agtctatagg cccacgggat tcgaacatcg attgaattcc gcgggcccgg gatccaccgg
      841 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      901 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      961 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1021 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1081 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1141 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1201 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1261 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1321 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1381 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1441 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1501 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1561 acaagtaaag cggccgctct agaggatcca agcttgcctc gagcagcgct gctcgagaga
     1621 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     1681 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     1741 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     1801 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     1861 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     1921 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     1981 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     2041 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     2101 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     2161 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     2221 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2281 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     2341 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2401 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     2461 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2521 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2581 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2641 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     2701 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     2761 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     2821 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     2881 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2941 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3001 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3061 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3121 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3181 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3241 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3301 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3361 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3421 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3481 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3541 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3601 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3661 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3721 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3781 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3841 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3901 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3961 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4021 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4081 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4141 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4861 tggccttttg c
//
