LOCUS       sequence-202477-        6286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP from EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6286)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 6286)
  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.EF1a.eGFP.WPRE.rBG
FEATURES             Location/Qualifiers
     source          1..6286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        98..1276
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          329..1267
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1224..1244
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          1390..1522
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1505..1524
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1567..1585
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1616..1625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1622..1624,1625..1627,1628..2341)
                     /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(1622..1624,1625..1627,1628..2338)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1667..1688)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1928..1947)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2275..2296
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2369..2957
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2422..2442)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2840..2851)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3017..3036)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3082..3137
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   3230..3359
                     /label=AAV2 ITR
     primer_bind     complement(3378..3395)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3378..3394)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3387..3409)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3536..3991
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3623..3642)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3833..3854
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4017..4121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4122..4190,4191..4982)
                     /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(4340..4359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5153..5741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5642..5661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6029..6050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6065..6070,6071..6088,6089..6095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6103..6119
                     /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     6108..6130
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6127..6143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6127..6143
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cttgtagtta atgattaacc cgccatgcta cttatctacc agggtaatgg ggatcctcta
       61 gaactatagc tagaattcta gtggatcccc ccgcgccggc tccggtgccc gtcagtgggc
      121 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
      181 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      241 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      301 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg
      361 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt
      421 gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta
      481 aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt
      541 gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta
      601 aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg
      661 ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc
      721 gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg
      781 ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc
      841 cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc
      901 gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc
      961 gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga
     1021 ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac
     1081 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt
     1141 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt
     1201 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc
     1261 atttcaggtg tcgtgagcta gagctttatt gcggtagttt atcacagtta aattgctaac
     1321 gcagtcagtg cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc
     1381 actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
     1441 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     1501 cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag
     1561 agtacttaat acgactcact ataggctagc ctcgagaatt gtacaattca cgcgggccgc
     1621 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1681 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1741 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1801 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1861 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1921 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1981 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2041 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2101 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2161 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2221 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2281 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2341 ataagctcgc gtgatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     2401 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2461 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2521 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2581 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2641 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2701 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2761 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2821 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2881 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2941 ttgggccgcc tccccgcatc gataccgtcg acccgggcgg cctcgaggac ggggtgaact
     3001 acgcctgagg atccgatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     3061 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     3121 attttttgtg tctctcactc ggaagcaatt cgttgatctg aatttcgacc acccataata
     3181 cccattaccc tggtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     3241 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3301 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3361 cttaattaac ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3421 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3481 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     3541 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3601 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3661 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3721 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3781 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3841 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3901 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3961 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4021 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4081 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4141 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4201 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4261 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4321 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4381 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4441 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4501 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4561 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4621 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4681 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4741 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4801 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4861 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4921 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4981 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5041 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5101 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5161 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5221 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5281 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5341 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5401 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5461 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5521 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5581 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5641 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5701 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5761 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5821 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5881 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     5941 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6001 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     6061 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     6121 ttcacacagg aaacagctat gaccatgatt acgccagatt taattaaggc cttaattagg
     6181 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcggcc tcagtgagcg
     6241 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttc
//
