LOCUS       sequence-390559-        6343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector plasmid expressing enhanced green fluorescent protein 
            (eGFP) under the cytomegalovirus (CMV) promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6343)
  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 6343)
  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-CMV-eGFP
FEATURES             Location/Qualifiers
     source          1..6343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     primer_bind     complement(189..208)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        310..689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        690..893
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     844..864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     890..914
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1012..1021
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1018..1020,1021..1023,1024..1737)
                     /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(1018..1020,1021..1023,1024..1734)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1063..1084)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1324..1343)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1671..1692
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1767..1783)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1768..1784)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    1877..2271
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     repeat_region   2626..2755
                     /label=AAV2 ITR
     rep_origin      3126..3639
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3213..3232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3423..3444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4121..4140)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4240..4262
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4300..4318)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4386..4490
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4491..4559,4560..5351)
                     /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(4709..4728)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5522..6110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6011..6030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctagcggcc tcggcctctg cataaataaa aaaaattagt cagccatgag cttggcccat
      241 tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac
      301 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      361 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      421 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      481 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      541 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      601 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      661 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      721 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      781 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      841 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      901 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      961 gggaccgatc cagcctcccc tcgaagctcg cgggcccggg atccaccggt cgccaccatg
     1021 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1081 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1141 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1201 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1261 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1321 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1381 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1441 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1501 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1561 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1621 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1681 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     1741 ggccgctcta gaggatccaa gcttatcgat accgtcgacc tcgagggccc agatctaatt
     1801 caccccacca gtgcaggctg cctatcagaa agtggtggct ggtgtggcta atgccctggc
     1861 ccacaagtat cactaagctc gctttcttgc tgtccaattt ctattaaagg ttcctttgtt
     1921 ccctaagtcc aactactaaa ctgggggata ttatgaaggg ccttgagcat ctggattctg
     1981 cctaataaaa aacatttatt ttcattgcaa tgatgtattt aaattatttc tgaatatttt
     2041 actaaaaagg gaatgtggga ggtcagtgca tttaaaacat aaagaaatga agagctagtt
     2101 caaaccttgg gaaaatacac tatatcttaa actccatgaa agaaggtgag gctgcaaaca
     2161 gctaatgcac attggcaaca gcccctgatg cctatgcctt attcatccct cagaaaagga
     2221 ttcaagtaga ggcttgattt ggaggttaaa gttttgctat gctgtatttt acattactta
     2281 ttgttttagc tgtcctcatg aatgtctttt cactacccat ttgcttatcc tgcatctctc
     2341 agccttgact ccactcagtt ctcttgctta gagataccac ctttcccctg aagtgttcct
     2401 tccatgtttt acggcgagat ggtttctcct cgcctggcca ctcagcctta gttgtctctg
     2461 ttgtcttata gaggtctact tgaagaagga aaaacagggg gcatggtttg actgtcctgt
     2521 gagcccttct tccctgcctc ccccactcac agtgacccgg aatccctcga catctagagc
     2581 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
     2641 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2701 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctggc
     2761 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2821 aatggcgatt ccgttgcaat ggctggcggt aatattgttc tggatattac cagcaaggcc
     2881 gatagtttga gttcttctac tcaggcaagt gatgttatta ctaatcaaag aagtattgcg
     2941 acaacggtta atttgcgtga tggacagact cttttactcg gtggcctcac tgattataaa
     3001 aacacttctc aggattctgg cgtaccgttc ctgtctaaaa tccctttaat cggcctcctg
     3061 tttagctccc gctctgattc taacgaggaa agcacgttat acgtgctcgt caaagcaacc
     3121 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3181 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3241 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3301 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3361 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3421 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3481 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3541 atttaacgcg aattttaaca aaatattaac gcttacaatt taaatatttg cttatacaat
     3601 cttcctgttt ttggggcttt tctgattatc aaccggggta catatgattg acatgctagt
     3661 tttacgatta ccgttcatcg attctcttgt ttgctccaga ctctcaggca atgacctgat
     3721 agcctttgta gagacctctc aaaaatagct accctctccg gcatgaattt atcagctaga
     3781 acggttgaat atcatattga tggtgatttg actgtctccg gcctttctca cccgtttgaa
     3841 tctttaccta cacattactc aggcattgca tttaaaatat atgagggttc taaaaatttt
     3901 tatccttgcg ttgaaataaa ggcttctccc gcaaaagtat tacagggtca taatgttttt
     3961 ggtacaaccg atttagcttt atgctctgag gctttattgc ttaattttgc taattctttg
     4021 ccttgcctgt atgatttatt ggatgttgga atcgcctgat gcggtatttt ctccttacgc
     4081 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     4141 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4201 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4261 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4321 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4381 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4441 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4501 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4561 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4621 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4681 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4741 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4801 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4861 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4921 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4981 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5041 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5101 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5161 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5221 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5281 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5341 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5401 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5461 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5521 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5581 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5641 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5701 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5761 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5821 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5881 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5941 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6001 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6061 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6121 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6181 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6241 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgccmaa
     6301 tmcgcaaacc gcctctcccc gcgcgttggc cgattcatta atg
//
