LOCUS       sequence-379932-        5271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector plasmid expressing enhanced green fluorescent protein 
            (eGFP) under the mouse phosphoglycerate kinase (mPGK) promoter that 
            is upstream of a beta-globin intron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5271)
  AUTHORS   Nieuwenhuis B, Haenzi B, Hilton S, Carnicer-Lombarte A, Hobo B, 
            Verhaagen J, Fawcett JW
  TITLE     Optimization of adeno-associated viral vector-mediated transduction 
            of the corticospinal tract: comparison of four promoters.
  JOURNAL   Gene Ther. 2020 Jun 23. pii: 10.1038/s41434-020-0169-1. doi: 
            10.1038/s41434-020-0169-1.
  PUBMED    32576975
REFERENCE   2  (bases 1 to 5271)
  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. 
            2020 Jun 23. pii: 10.1038/s41434-020-0169-1. doi: 
            10.1038/s41434-020-0169-1."
COMMENT     Sequence Label: AAV-mPGK-eGFP (beta-globin intron)
FEATURES             Location/Qualifiers
     source          1..5271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   8..137
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        187..686
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(222..245)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     619..638
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     intron          763..1141
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     regulatory      1242..1251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1248..1250,1251..1253,1254..1967)
                     /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(1248..1250,1251..1253,1254..1964)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1293..1314)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1554..1573)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1901..1922
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2025..2501
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2224..2243)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2541..2681
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2541..2670
                     /label=AAV2 ITR
     rep_origin      2756..3211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2843..2862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3053..3074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3228..3247)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3347..3369
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3407..3425)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3493..3597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3598..3666,3667..4458)
                     /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(3816..3835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4629..5217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5118..5137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
       61 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
      121 catcactagg ggttcctgcg gccgcacgcg cgatcacgag actagcctcg acgatggtcg
      181 agtaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
      241 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
      301 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
      361 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
      421 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
      481 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
      541 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
      601 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
      661 cctcttcctc atctccgggc ctttcgacct ctagcgggat cggattcgaa tcccggccgg
      721 gaacggtgca ttggaacgcg gattccccgt gccaagagtg acgtaagtac cgcctataga
      781 gtctataggc ccacaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt
      841 ctaatacttt ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct
      901 ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct
      961 gcatataaat atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata
     1021 gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag gctggattat
     1081 tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct tcctcccaca
     1141 gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag aattgggatt
     1201 cgaacatcga ttgaattccg cgggcccggg atccaccggt cgccaccatg gtgagcaagg
     1261 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1321 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1381 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1441 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     1501 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1561 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1621 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1681 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     1741 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1801 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1861 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1921 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgctcta
     1981 gaggatccaa gcttgcctcg agcagcgctg ctcgagagat ctacgggtgg catccctgtg
     2041 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     2101 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     2161 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     2221 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     2281 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     2341 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     2401 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     2461 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     2521 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2581 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2641 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2701 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2821 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2881 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2941 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3061 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3121 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3181 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3241 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3301 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3361 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3421 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4141 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4741 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc t
//
