LOCUS       sequence-485240-        4566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid expressing EGFP, control for plasmid 242957.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4566)
  AUTHORS   Addgene Research Program
  TITLE     Addgene AAV Transfer Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4566)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAG AAV CMV EGFP
FEATURES             Location/Qualifiers
     source          1..4566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..122
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     114..131
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     115..131
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     141..160
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        141..159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     183..199
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     184..200
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   210..350
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   221..350
                     /label=AAV2 ITR
     promoter        442..644
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     596..616
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(705..707,708..710,711..1421)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(705..707,708..710,711..1421)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(750..771)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1011..1030)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1358..1379
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1514..1635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1551..1570)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1605..1624
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1662..1802
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1662..1791
                     /label=AAV2 ITR
     primer_bind     complement(1893..1913)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1893..1911)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1932..1948)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1932..1948)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1945..1967)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1956..1972
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1980..1986,1987..2004,2005..2010))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2025..2046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2334..2922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2414..2433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3093..3884,3885..3953))
                     /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     3716..3735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3954..4058)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4084..4539)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4221..4242)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4433..4452
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
       61 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      121 cgacggccag tgagcgcgcg taatacgact cactataggg cgaatgggta ccggaccccc
      181 cctcgaggtc gacggtatcg ggggcatgcc ctgcaggcag ctgcgcgctc gctcgctcac
      241 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
      301 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcag
      361 ccgtgtttta gagctagaaa tagcaagtta aaataaggct agtccgtagc gcgtgcgcca
      421 attctgcaga caaattctag agtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      481 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      541 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      601 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagcggtcac cggtttagtg
      661 aaccgtcaga tccgctagcg ctaccggtcg ccaccatgtt cgaaatggtg agcaagggcg
      721 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      781 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      841 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      901 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      961 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1021 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1081 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1141 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1201 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1261 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1321 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1381 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggatcctga acgcgtgtag
     1441 tagttcaagt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     1501 tgagaggtgt acaaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1561 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1621 catcaatgta tcttagttaa cacgtgcgac cgagcggccg caggaacccc tagtgatgga
     1681 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     1741 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     1801 ggggcgccaa gccgaattct gcagatatcc atcacactgg cggccgctcg actagagcgg
     1861 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     1921 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1981 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2041 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2101 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2161 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2221 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2281 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2341 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2401 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2461 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2521 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2581 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2641 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2701 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2761 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2821 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2881 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2941 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3001 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3061 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3121 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3181 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3241 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3301 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3361 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3421 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3481 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3541 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3601 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3661 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3721 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3781 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3841 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3901 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3961 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4021 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4081 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4141 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4201 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4261 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4321 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4381 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4441 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4501 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     4561 gcaact
//
