LOCUS       sequence-201073-        7496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of GFP-Cre from CamKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7496)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 7496)
  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.CamKII.HI.GFP-Cre.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..7496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     regulatory      1673..1682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1679..1681,1682..1684,1685..2395)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1679..1681,1682..1684,1685..2395)
                     /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(1985..2004)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2332..2353
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2402..2431
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2438..2458
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2459..3487
                     /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    3508..4096
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3561..3581)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3979..3990)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4129..4263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4166..4185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4220..4239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4292..4421
                     /label=AAV2 ITR
     primer_bind     complement(4440..4457)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4440..4456)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4449..4471)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4596..5051
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4683..4702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4893..4914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5068..5087)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5187..5209
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5247..5265)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5333..5437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5438..5506,5507..6298)
                     /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(5656..5675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6469..7057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6958..6977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7345..7366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7381..7386,7387..7404,7405..7411)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7419..7435
                     /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     7424..7446
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7443..7459
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7443..7459
                     /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 gcggccgcac gcgtactagg tagccatgct ctaggaagat ctaacattat
      181 ggccttaggt cacttcatct ccatggggtt cttcttctga ttttctagaa aatgagatgg
      241 gggtgcagag agcttcctca gtgacctgcc cagggtcaca tcagaaatgt cagagctaga
      301 acttgaactc agattactaa tcttaaattc catgccttgg gggcatgcaa gtacgatata
      361 cagaaggagt gaactcatta gggcagatga ccaatgagtt taggaaagaa gagtccaggg
      421 cagggtacat ctacaccacc cgcccagccc tgggtgagtc cagccacgtt cacctcatta
      481 tagttgcctc tctccagtcc taccttgacg ggaagcacaa gcagaaactg ggacaggagc
      541 cccaggagac caaatcttca tggtccctct gggaggatgg gtggggagag ctgtggcaga
      601 ggcctcagga ggggccctgc tgctcagtgg tgacagatag gggtgagaaa gcagacagag
      661 tcattccgtc agcattctgg gtctgtttgg tacttcttct cacgctaagg tggcggtgtg
      721 atatgcacaa tggctaaaaa gcagggagag ctggaaagaa acaaggacag agacagaggc
      781 caagtcaacc agaccaattc ccagaggaag caaagaaacc attacagaga ctacaagggg
      841 gaagggaagg agagatgaat tagcttcccc tgtaaacctt agaacccagc tgttgccagg
      901 gcaacggggc aatacctgtc tcttcagagg agatgaagtt gccagggtaa ctacatcctg
      961 tctttctcaa ggaccatccc agaatgtggc acccactagc cgttaccata gcaactgcct
     1021 ctttgcccca cttaatccca tcccgtctgt taaaagggcc ctatagttgg aggtggggga
     1081 ggtaggaaga gcgatgatca cttgtggact aagtttgttc gcatcccctt ctccaacccc
     1141 ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggcccaca cagtcctgca
     1201 gtattgtgta tataaggcca gggcaaagag gagcaggttt taaagtgaaa ggcaggcagg
     1261 tgttggggag gcagttaccg gggcaacggg aacagggcgt ttcggaggtg gttgccatgg
     1321 ggacctggat gctgacgaag gctcgcggga ccgatccagc ctccgtaccg gttcgaacag
     1381 gtaagcgccc ctaaaatccc tttgggcaca atgtgtcctg aggggagagg cagcgacctg
     1441 tagatgggac gggggcacta accctcaggt ttggggcttc tgaatgtgag tatcgccatg
     1501 taagcccagt atttggccaa tctcagaaag ctcctggtcc ctggagggat ggagagagaa
     1561 aaacaaacag ctcctggagc agggagagtg ctggcctctt gctctccggc tccctctgtt
     1621 gccctctggt ttctccccag gttcgaatct agaggatccc cgggtaccgg tcgccaccat
     1681 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1741 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1801 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1861 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1921 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1981 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2041 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2101 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2161 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2221 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2281 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2341 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     2401 agagcaaaag ctgatttctg aggaggatct gggaggaccc aagaagaaga ggaaggtgtc
     2461 caatttactg accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa cgagtgatga
     2521 ggttcgcaag aacctgatgg acatgttcag ggatcgccag gcgttttctg agcatacctg
     2581 gaaaatgctt ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga ataaccggaa
     2641 atggtttccc gcagaacctg aagatgttcg cgattatctt ctatatcttc aggcgcgcgg
     2701 tctggcagta aaaactatcc agcaacattt gggccagcta aacatgcttc atcgtcggtc
     2761 cgggctgcca cgaccaagtg acagcaatgc tgtttcactg gttatgcggc ggatccgaaa
     2821 agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac gcactgattt
     2881 cgaccaggtt cgttcactca tggaaaatag cgatcgctgc caggatatac gtaatctggc
     2941 atttctgggg attgcttata acaccctgtt acgtatagcc gaaattgcca ggatcagggt
     3001 taaagatatc tcacgtactg acggtgggag aatgttaatc catattggca gaacgaaaac
     3061 gctggttagc accgcaggtg tagagaaggc acttagcctg ggggtaacta aactggtcga
     3121 gcgatggatt tccgtctctg gtgtagctga tgatccgaat aactacctgt tttgccgggt
     3181 cagaaaaaat ggtgttgccg cgccatctgc caccagccag ctatcaactc gcgccctgga
     3241 agggattttt gaagcaactc atcgattgat ttacggcgct aaggatgact ctggtcagag
     3301 atacctggcc tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc
     3361 tggagtttca ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtcat
     3421 gaactatatc cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg
     3481 cgattagctc gaatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     3541 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3601 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3661 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3721 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3781 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3841 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3901 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     3961 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4021 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4081 tgggccgcct ccccgcatcg ataccgtcga tcgagaaatc gatcgcccaa cttgtttatt
     4141 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4201 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4261 atctccggac acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     4321 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4381 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca
     4441 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4501 cttgcagcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4561 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4621 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     4681 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4741 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4801 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4861 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4921 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4981 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5041 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5101 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5161 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5221 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5281 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5341 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5401 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5461 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5521 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5581 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5641 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5701 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5761 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5821 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5881 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5941 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6001 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6061 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6121 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6181 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6241 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6301 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6361 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6421 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6601 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7081 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7141 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7201 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7261 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     7321 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     7381 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7441 cacaggaaac agctatgacc atgattacgc cagatttaat taaggcctta attagg
//
