LOCUS       sequence-202462-        6464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP-Cre from CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6464)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 6464)
  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: pAAV.CMV.HI.eGFP-Cre.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   88..217
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        294..597
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        598..801
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     752..772
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     798..822
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1195..1204
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1201..1203,1204..1206,1207..1917)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1201..1203,1204..1206,1207..1917)
                     /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(1507..1526)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1854..1875
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1924..1953
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1960..1980
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1981..3009
                     /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    3030..3618
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3083..3103)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3501..3512)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3651..3785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3688..3707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3742..3761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3814..3954
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3814..3943
                     /label=AAV2 ITR
     rep_origin      4029..4484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4116..4135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4326..4347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4501..4520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4620..4642
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4680..4698)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4766..4870
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4871..4939,4940..5731)
                     /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(5089..5108)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5902..6464,1..26)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6391..6410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
       61 ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg
      121 aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
      181 agagagggag tggccaactc catcactagg ggttcctgcg gccgcacgcg tactagttat
      241 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      301 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      361 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      421 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      481 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      541 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      601 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      661 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      721 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      781 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
      841 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccgtac cggttcgaac
      901 aggtaagcgc ccctaaaatc cctttgggca caatgtgtcc tgaggggaga ggcagcgacc
      961 tgtagatggg acgggggcac taaccctcag gtttggggct tctgaatgtg agtatcgcca
     1021 tgtaagccca gtatttggcc aatctcagaa agctcctggt ccctggaggg atggagagag
     1081 aaaaacaaac agctcctgga gcagggagag tgctggcctc ttgctctccg gctccctctg
     1141 ttgccctctg gtttctcccc aggttcgaat ctagaggatc cccgggtacc ggtcgccacc
     1201 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1261 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1321 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1381 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1441 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1501 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1561 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1621 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1681 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1741 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1801 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1861 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
     1921 ggagagcaaa agctgatttc tgaggaggat ctgggaggac ccaagaagaa gaggaaggtg
     1981 tccaatttac tgaccgtaca ccaaaatttg cctgcattac cggtcgatgc aacgagtgat
     2041 gaggttcgca agaacctgat ggacatgttc agggatcgcc aggcgttttc tgagcatacc
     2101 tggaaaatgc ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg
     2161 aaatggtttc ccgcagaacc tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc
     2221 ggtctggcag taaaaactat ccagcaacat ttgggccagc taaacatgct tcatcgtcgg
     2281 tccgggctgc cacgaccaag tgacagcaat gctgtttcac tggttatgcg gcggatccga
     2341 aaagaaaacg ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga acgcactgat
     2401 ttcgaccagg ttcgttcact catggaaaat agcgatcgct gccaggatat acgtaatctg
     2461 gcatttctgg ggattgctta taacaccctg ttacgtatag ccgaaattgc caggatcagg
     2521 gttaaagata tctcacgtac tgacggtggg agaatgttaa tccatattgg cagaacgaaa
     2581 acgctggtta gcaccgcagg tgtagagaag gcacttagcc tgggggtaac taaactggtc
     2641 gagcgatgga tttccgtctc tggtgtagct gatgatccga ataactacct gttttgccgg
     2701 gtcagaaaaa atggtgttgc cgcgccatct gccaccagcc agctatcaac tcgcgccctg
     2761 gaagggattt ttgaagcaac tcatcgattg atttacggcg ctaaggatga ctctggtcag
     2821 agatacctgg cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc
     2881 gctggagttt caataccgga gatcatgcaa gctggtggct ggaccaatgt aaatattgtc
     2941 atgaactata tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct gctggaagat
     3001 ggcgattagc tcgaatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3061 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3121 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3181 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3241 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3301 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3361 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3421 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3481 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3541 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3601 tttgggccgc ctccccgcat cgataccgtc gatcgagaaa tcgatcgccc aacttgttta
     3661 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3721 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3781 ggatctccgg acacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3841 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3901 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3961 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4021 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4081 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4141 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4201 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4261 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4321 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4381 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4441 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4501 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4561 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4621 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4681 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4741 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4801 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4861 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4921 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4981 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5041 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5101 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5161 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5221 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5281 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5341 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5401 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5461 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5521 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5581 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5641 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5701 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5761 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5821 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5881 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5941 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6001 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6061 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6121 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6181 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6241 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6301 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6361 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6421 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgct
//
