LOCUS       sequence-202384-        5193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP from CamKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5193)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5193)
  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: pENN.AAV.CamKII0.4.eGFP.WPRE.rBG
FEATURES             Location/Qualifiers
     source          1..5193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(607..609,610..612,613..1326)
                     /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(607..609,610..612,613..1323)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(652..673)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(913..932)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1260..1281
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1393..1981
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1446..1466)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1864..1875)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2030..2049)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2095..2150
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2243..2372
                     /label=AAV2 ITR
     primer_bind     complement(2391..2408)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2391..2407)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2400..2422)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2549..3004
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2636..2655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2846..2867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3030..3134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3135..3203,3204..3995)
                     /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(3353..3372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4166..4754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4655..4674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5042..5063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5078..5083,5084..5101,5102..5108)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5116..5132
                     /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)."
     primer_bind     5121..5143
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5140..5156
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5140..5156
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgkcg kscgrscwww
       61 gsycgsscgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gcatgcctaa cttgtggact aagtttgttc acatcccctt
      241 ctccaacccc ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggcccaca
      301 cagtcctgca gtattgtgta tataaggcca gggcaaagag gagcaggttt taaagtgaaa
      361 ggcaggcagg tgttggggag gcagttaccg gggcaacggg aacagggcgt ttcggaggtg
      421 gttgccatgg ggacctggat gctgacgaag gctcgcgagg ctgtgagcag ccacagtgcc
      481 ctgctcagaa gccccaagct cgtcagtcaa gccggttctc cgtttgcact caggagcacg
      541 ggcaggcgag tggcccctag ttctgggggc agctctagag ccgatccccg ggtaccggtc
      601 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      661 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      721 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      781 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      841 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      901 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      961 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1021 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1081 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1141 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1201 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1261 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1321 aagtaaagcg gccgcgactc tagaattgta cctctagagt cgacccgggc ggcctcgatc
     1381 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     1441 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     1501 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     1561 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     1621 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     1681 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     1741 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     1801 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     1861 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     1921 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     1981 catcgatacc gtcgatcgag gacggggtga actacgcctg aggatccgat ctttttccct
     2041 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     2101 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagca
     2161 attcgttgat ctgaatttcg accacccata atacccatta ccctggtaga taagtagcat
     2221 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     2281 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2341 cgggcggcct cagtgagcga gcgagcgcgc agccttaatt aacctaattc actggccgtc
     2401 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2461 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2521 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     2581 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2641 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2701 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2761 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2821 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2881 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2941 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3001 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3061 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3121 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3181 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3241 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3301 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3361 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3421 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3481 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3541 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3601 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3661 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3721 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3781 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3841 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3901 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3961 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4021 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4081 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4801 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4861 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4921 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4981 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5041 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5101 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5161 attacgccag atttaattaa ggccttaatt agg
//
