LOCUS       sequence-72100-e        8002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adeno-associated virus (AAV) encoding a green fluorescent 
            protein/Cre recombinase (GFP/Cre) fusion protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8002)
  AUTHORS   Kaspar BK, Vissel B, Bengoechea T, Crone S, Randolph-Moore L, Muller
            R, Brandon EP, Schaffer D, Verma IM, Lee KF, Heinemann SF, Gage FH
  TITLE     Adeno-associated virus effectively mediates conditional gene 
            modification in the brain.
  JOURNAL   Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):2320-5. Epub 2002 Feb 
            12.
  PUBMED    11842206
REFERENCE   2  (bases 1 to 8002)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2002-02-19"; volume: "99(4)"; pages: 
            "2320-5. Epub 2002 Feb 12"
COMMENT     Sequence Label: AAV-GFP/Cre
FEATURES             Location/Qualifiers
     source          1..8002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     primer_bind     complement(198..217)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        319..698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        699..902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     853..873
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     899..923
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1036..1511
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      1585..1594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1591..1593,1594..1596,1597..2307)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1591..1593,1594..1596,1597..2307)
                     /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(1897..1916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2244..2265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2314..2343
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2350..2370
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2371..3399
                     /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"
     polyA_signal    3487..3881
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      4786..5299
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4873..4892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5083..5104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5780..5799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5899..5921
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5959..5977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6045..6149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6150..6218,6219..7010)
                     /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(6368..6387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7181..7769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7670..7689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctagag gatccggcct cggcctctgc ataaataaaa aaaattagtc agccatgagc
      241 ttggcccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
      301 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      361 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggactattta cggtaaactg
      541 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      721 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      781 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      841 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      901 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
      961 gaagacaccg ggaccgatcc agcctcccct cgaagcttac atgtggtacc gagctcggat
     1021 cctgagaact tcagggtgag tctatgggac ccttgatgtt ttctttcccc ttcttttcta
     1081 tggttaagtt catgtcatag gaaggggaga agtaacaggg tacacatatt gaccaaatca
     1141 gggtaatttt gcatttgtaa ttttaaaaaa tgctttcttc ttttaatata cttttttgtt
     1201 tatcttattt ctaatacttt ccctaatctc tttctttcag ggcaataatg atacaatgta
     1261 tcatgcctct ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaatag
     1321 caatatttct gcatataaat atttctgcat ataaattgta actgatgtaa gaggtttcat
     1381 attgctaata gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag
     1441 gctggattat tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct
     1501 tcctcccaca gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag
     1561 cacgtgaatt cgatgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     1621 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1681 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1741 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1801 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1861 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1921 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1981 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2041 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2101 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2161 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2221 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2281 actctcggca tggacgagct gtacaagtcc ggagagcaaa agctgatttc tgaggaggat
     2341 ctgggaggac ccaagaagaa gaggaaggtg tccaatttac tgaccgtaca ccaaaatttg
     2401 cctgcattac cggtcgatgc aacgagtgat gaggttcgca agaacctgat ggacatgttc
     2461 agggatcgcc aggcgttttc tgagcatacc tggaaaatgc ttctgtccgt ttgccggtcg
     2521 tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc ccgcagaacc tgaagatgtt
     2581 cgcgattatc ttctatatct tcaggcgcgc ggtctggcag taaaaactat ccagcaacat
     2641 ttgggccagc taaacatgct tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat
     2701 gctgtttcac tggttatgcg gcggatccga aaagaaaacg ttgatgccgg tgaacgtgca
     2761 aaacaggctc tagcgttcga acgcactgat ttcgaccagg ttcgttcact catggaaaat
     2821 agcgatcgct gccaggatat acgtaatctg gcatttctgg ggattgctta taacaccctg
     2881 ttacgtatag ccgaaattgc caggatcagg gttaaagata tctcacgtac tgacggtggg
     2941 agaatgttaa tccatattgg cagaacgaaa acgctggtta gcaccgcagg tgtagagaag
     3001 gcacttagcc tgggggtaac taaactggtc gagcgatgga tttccgtctc tggtgtagct
     3061 gatgatccga ataactacct gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct
     3121 gccaccagcc agctatcaac tcgcgccctg gaagggattt ttgaagcaac tcatcgattg
     3181 atttacggcg ctaaggatga ctctggtcag agatacctgg cctggtctgg acacagtgcc
     3241 cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt caataccgga gatcatgcaa
     3301 gctggtggct ggaccaatgt aaatattgtc atgaactata tccgtaacct ggatagtgaa
     3361 acaggggcaa tggtgcgcct gctggaagat ggcgattagc tcgagcaatt caccccacca
     3421 gtgcaggctg cctatcagaa agtggtggct ggtgtggcta atgccctggc ccacaagtat
     3481 cactaagctc gctttcttgc tgtccaattt ctattaaagg ttcctttgtt ccctaagtcc
     3541 aactactaaa ctgggggata ttatgaaggg ccttgagcat ctggattctg cctaataaaa
     3601 aacatttatt ttcattgcaa tgatgtattt aaattatttc tgaatatttt actaaaaagg
     3661 gaatgtggga ggtcagtgca tttaaaacat aaagaaatga agagctagtt caaaccttgg
     3721 gaaaatacac tatatcttaa actccatgaa agaaggtgag gctgcaaaca gctaatgcac
     3781 attggcaaca gcccctgatg cctatgcctt attcatccct cagaaaagga ttcaagtaga
     3841 ggcttgattt ggaggttaaa gttttgctat gctgtatttt acattactta ttgatttagc
     3901 tgcctcatga atgtcttttc actacccatt tgcttatcct gcatctctca gccttgactc
     3961 cactcagttc tcttgcttag agataccacc tttcccctga agtgttcctt ccatgtttta
     4021 cggcgagatg gtttctcctc gcctggccac tcagcttagt tgtctctgtt gtcttataga
     4081 ggtctacttg aagaaggaaa aacagggggc atggtttgac tgtcctgtga gcccttcttc
     4141 ctgcctcccc cactcacagt gacccggaat ccctcgacat ggcagtctag agcatggcta
     4201 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4261 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4321 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgccagc tggcgtaata
     4381 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     4441 attccagacg attgagcgtc aaaatgtagg tatttccatg agcgtttttc ctgttgcaat
     4501 ggctggcggt aatattgttc tggatattac cagcaaggcc gatagtttga gttcttctac
     4561 tcaggcaagt gatgttatta ctaatcaaag aagtattgcg acaacggtta atttgcgtga
     4621 tggacagact cttttactcg gtggcctcac tgattataaa aacacttctc aggattctgg
     4681 cgtaccgttc ctgtctaaaa tccctttaat cggcctcctg tttagctccc gctctgattc
     4741 taacgaggaa agcacgttat acgtgctcgt caaagcaacc atagtacgcg ccctgtagcg
     4801 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4861 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4921 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4981 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5041 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5101 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5161 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5221 aaatattaac gtttacaatt taaatatttg cttatacaat cttcctgttt ttggggcttt
     5281 tctgattatc aaccggggta catatgattg acatgctagt tttacgatta ccgttcatcg
     5341 attctcttgt ttgctccaga ctctcaggca atgacctgat agcctttgta gagacctctc
     5401 aaaaatagct accctctccg gcatgaattt atcagctaga acggttgaat atcatattga
     5461 tggtgatttg actgtctccg gcctttctca cccgtttgaa tctttaccta cacattactc
     5521 aggcattgca tttaaaatat atgagggttc taaaaatttt tatccttgcg ttgaaataaa
     5581 ggcttctccc gcaaaagtat tacagggtca taatgttttt ggtacaaccg atttagcttt
     5641 atgctctgag gctttattgc ttaattttgc taattctttg ccttgcctgt atgatttatt
     5701 ggatgttgga attcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5761 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5821 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5881 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5941 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     6001 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6121 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6241 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6301 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6361 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6421 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6481 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6541 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6601 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6661 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6721 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6781 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6841 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6901 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6961 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7021 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7081 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7141 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7201 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7261 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7321 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7381 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7441 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7501 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7561 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7621 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7681 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7741 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7801 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7861 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7921 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7981 cgcgttggcc gattcattaa tg
//
