LOCUS       sequence-201078-        5687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP from tMCK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5687)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5687)
  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.tMCK.PI.eGFP.WPRE.bGH
FEATURES             Location/Qualifiers
     source          1..5687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          1015..1147
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1130..1149
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1159..1168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1165..1167,1168..1170,1171..1884)
                     /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(1165..1167,1168..1170,1171..1881)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1210..1231)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1471..1490)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1818..1839
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1951..1971)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2369..2380)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2447..2464)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2453..2660
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2748..2877
                     /label=AAV2 ITR
     primer_bind     complement(2896..2913)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2896..2912)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2905..2927)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3054..3509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3141..3160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3351..3372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3535..3639
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3640..3708,3709..4500)
                     /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(3858..3877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4671..5259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5160..5179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5547..5568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5583..5588,5589..5606,5607..5613)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5621..5637
                     /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     5626..5648
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5645..5661
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5645..5661
                     /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 tatctacgta gccatgctct
      181 aggaagatcg gaattcgccc ttaagctagc cactacgggt ctaggctgcc catgtaagga
      241 ggcaaggcct ggggacaccc gagatgcctg gttataatta accccaacac ctgctgcccc
      301 ccccccccca acacctgctg cctgagcctg agcggttacc ccaccccggt gcctgggtct
      361 taggctctgt acaccatgga ggagaagctc gctctaaaaa taaccctgtc cctggtgggc
      421 ccactacggg tctaggctgc ccatgtaagg aggcaaggcc tggggacacc cgagatgcct
      481 ggttataatt aaccccaaca cctgctgccc cccccccccc aacacctgct gcctgagcct
      541 gagcggttac cccaccccgg tgcctgggtc ttaggctctg tacaccatgg aggagaagct
      601 cgctctaaaa ataaccctgt ccctggtggg ccactacggg tctaggctgc ccatgtaagg
      661 aggcaaggcc tggggacacc cgagatgcct ggttataatt aaccccaaca cctgctgccc
      721 cccccccccc aacacctgct gcctgagcct gagcggttac cccaccccgg tgcctgggtc
      781 ttaggctctg tacaccatgg aggagaagct cgctctaaaa ataaccctgt ccctggtggg
      841 cccctccctg gggacagccc ctcctggcta gtcacaccct gtaggctcct ctatataacc
      901 caggggcaca ggggctgccc ccgggtcacg gggatcctct agacctccga acggtaagag
      961 cctagcatgt agaactggtt accctgcaga agttggtcgt gaggcactgg gcaggtaagt
     1021 atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga
     1081 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
     1141 tccacaggtg tccaggcggc cgccatggtg agcaagggcg aggagctgtt caccggggtg
     1201 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1261 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1321 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1381 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1441 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1501 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1561 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1621 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1681 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1741 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1801 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1861 ggcatggacg agctgtacaa gtaataagct tggatccaat caacctctgg attacaaaat
     1921 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1981 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2041 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2101 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2161 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2221 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2281 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     2341 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2401 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcga gatctgcctc gactgtgcct
     2461 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2521 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2581 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2641 aatagcaggc atgctgggga ctcgagttaa gggcgaattc ccgataagga tcttcctaga
     2701 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     2761 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     2821 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagcct
     2881 taattaacct aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2941 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3001 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     3061 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3121 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3181 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3241 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3301 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3361 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3421 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3481 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     3541 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3601 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3661 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3721 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3781 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3841 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3901 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3961 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4021 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4081 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4141 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4201 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4261 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4321 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4381 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4441 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4501 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4561 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4621 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4681 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4741 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4801 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4861 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4921 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4981 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5041 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5101 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5161 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5221 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5281 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5341 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5401 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     5461 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5521 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     5581 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     5641 cacacaggaa acagctatga ccatgattac gccagattta attaagg
//
