LOCUS       sequence-201069-        6308 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 6308)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 6308)
  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..6308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        226..1404
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          457..1395
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1352..1372
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          1518..1650
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1633..1652
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1695..1713
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1744..1753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1750..1752,1753..1755,1756..2469)
                     /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(1750..1752,1753..1755,1756..2466)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1795..1816)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2056..2075)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2403..2424
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2497..3085
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2550..2570)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2968..2979)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3145..3164)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3210..3265
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   3358..3487
                     /label=AAV2 ITR
     primer_bind     complement(3506..3523)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3506..3522)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3515..3537)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3664..4119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3751..3770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3961..3982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4145..4249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4250..4318,4319..5110)
                     /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(4468..4487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5281..5869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5770..5789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6157..6178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6193..6198,6199..6216,6217..6223)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6231..6247
                     /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     6236..6258
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6255..6271
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6255..6271
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gaattctagt ggatcccccc gcgccggctc cggtgcccgt
      241 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
      301 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
      361 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
      421 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
      481 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
      541 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
      601 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
      661 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
      721 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
      781 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
      841 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
      901 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
      961 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     1021 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
     1081 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     1141 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     1201 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     1261 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     1321 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     1381 tttcttccat ttcaggtgtc gtgagctaga gctttattgc ggtagtttat cacagttaaa
     1441 ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agaagttggt
     1501 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
     1561 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
     1621 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt
     1681 aaggctagag tacttaatac gactcactat aggctagcct cgagaattgt acaattcacg
     1741 cgggccgcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1801 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1861 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1921 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1981 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2041 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2101 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2161 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2221 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2281 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2341 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2401 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2461 tacaagtaat aagctcgcgt gatcaagctt atcgataatc aacctctgga ttacaaaatt
     2521 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2581 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2641 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2701 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2761 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2821 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2881 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     2941 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3001 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3061 atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc tcgaggacgg
     3121 ggtgaactac gcctgaggat ccgatctttt tccctctgcc aaaaattatg gggacatcat
     3181 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     3241 gtgttggaat tttttgtgtc tctcactcgg aagcaattcg ttgatctgaa tttcgaccac
     3301 ccataatacc cattaccctg gtagataagt agcatggcgg gttaatcatt aactacaagg
     3361 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3421 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3481 cgcgcagcct taattaacct aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3541 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3601 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     3661 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3721 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3781 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3841 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3901 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3961 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4021 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4081 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     4141 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4201 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4261 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4321 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4381 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4441 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4501 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4561 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4621 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4681 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4741 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4801 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4861 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4921 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4981 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5041 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5101 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5161 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5221 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5281 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5341 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5401 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5461 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5521 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5581 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5641 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5701 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5761 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5821 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5881 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5941 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6001 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     6061 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6121 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     6181 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6241 ataacaattt cacacaggaa acagctatga ccatgattac gccagattta attaaggcct
     6301 taattagg
//
