LOCUS       sequence-201061-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP-Cre from hSyn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 6733)
  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.hSyn.HI.eGFP-Cre.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        234..681
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1038..1047
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1044..1046,1047..1049,1050..1760)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1044..1046,1047..1049,1050..1760)
                     /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(1350..1369)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1697..1718
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1767..1796
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1803..1823
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1824..2852
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    2872..3460
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2925..2945)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3343..3354)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3500..3621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3511..3530)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3565..3584
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3783..3912
                     /label=AAV2 ITR
     primer_bind     complement(3931..3948)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3931..3947)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3940..3962)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4089..4544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4176..4195)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4386..4407
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4570..4674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4675..4743,4744..5535)
                     /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(4893..4912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5706..6294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6195..6214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6582..6603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6618..6623,6624..6641,6642..6648)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6656..6672
                     /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     6661..6683
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6680..6696
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6680..6696
                     /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 aggaagatcc tctagaacta tagctagcat gcctgcagag ggccctgcgt atgagtgcaa
      241 gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac
      301 ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg
      361 aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg
      421 ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg
      481 tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc
      541 gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat
      601 ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc
      661 gtgtcgtgcc tgagagcgca gtcgaattcg cgtggtacct ctagagtcga cgggaccgat
      721 ccagcctccg taccggttcg aacaggtaag cgcccctaaa atccctttgg gcacaatgtg
      781 tcctgagggg agaggcagcg acctgtagat gggacggggg cactaaccct caggtttggg
      841 gcttctgaat gtgagtatcg ccatgtaagc ccagtatttg gccaatctca gaaagctcct
      901 ggtccctgga gggatggaga gagaaaaaca aacagctcct ggagcaggga gagtgctggc
      961 ctcttgctct ccggctccct ctgttgccct ctggtttctc cccaggttcg aatctagagg
     1021 atccccgggt accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1081 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1141 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1201 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1261 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1321 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1381 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1441 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1501 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1561 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1621 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1681 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1741 gcatggacga gctgtacaag tccggagagc aaaagctgat ttctgaggag gatctgggag
     1801 gacccaagaa gaagaggaag gtgtccaatt tactgaccgt acaccaaaat ttgcctgcat
     1861 taccggtcga tgcaacgagt gatgaggttc gcaagaacct gatggacatg ttcagggatc
     1921 gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg
     1981 catggtgcaa gttgaataac cggaaatggt ttcccgcaga acctgaagat gttcgcgatt
     2041 atcttctata tcttcaggcg cgcggtctgg cagtaaaaac tatccagcaa catttgggcc
     2101 agctaaacat gcttcatcgt cggtccgggc tgccacgacc aagtgacagc aatgctgttt
     2161 cactggttat gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg
     2221 ctctagcgtt cgaacgcact gatttcgacc aggttcgttc actcatggaa aatagcgatc
     2281 gctgccagga tatacgtaat ctggcatttc tggggattgc ttataacacc ctgttacgta
     2341 tagccgaaat tgccaggatc agggttaaag atatctcacg tactgacggt gggagaatgt
     2401 taatccatat tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag aaggcactta
     2461 gcctgggggt aactaaactg gtcgagcgat ggatttccgt ctctggtgta gctgatgatc
     2521 cgaataacta cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca
     2581 gccagctatc aactcgcgcc ctggaaggga tttttgaagc aactcatcga ttgatttacg
     2641 gcgctaagga tgactctggt cagagatacc tggcctggtc tggacacagt gcccgtgtcg
     2701 gagccgcgcg agatatggcc cgcgctggag tttcaatacc ggagatcatg caagctggtg
     2761 gctggaccaa tgtaaatatt gtcatgaact atatccgtaa cctggatagt gaaacagggg
     2821 caatggtgcg cctgctggaa gatggcgatt agctcgatca agcttatcga taatcaacct
     2881 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2941 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3001 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3061 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3121 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3181 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3241 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     3301 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3361 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3421 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgatcgagg
     3481 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3541 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3601 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3661 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     3721 aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     3781 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3841 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3901 gcgagcgcgc agccttaatt aacctaattc actggccgtc gttttacaac gtcgtgactg
     3961 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4021 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4081 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4141 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4201 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4261 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4321 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4381 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4441 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4501 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     4561 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4621 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4681 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4741 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4801 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4861 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4921 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4981 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5041 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5101 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5161 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5221 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5281 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5341 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5401 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5461 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5521 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5581 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5641 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5701 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5761 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5821 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5881 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5941 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6001 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6061 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6121 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6181 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6241 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6301 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6361 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6421 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6481 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6541 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6601 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6661 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag atttaattaa
     6721 ggccttaatt agg
//
