LOCUS       sequence-182646-        7956 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 7956)
  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 7956)
  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..7956
                     /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"
     repeat_region   4242..4371
                     /label=AAV2 ITR
     rep_origin      4742..5255
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4829..4848)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5039..5060
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5737..5756)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5856..5878
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5916..5934)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6002..6106
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6107..6175,6176..6967)
                     /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(6325..6344)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7138..7726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7627..7646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcagctgc 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 ggagtattta 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 ttgttttagc
     3901 tgtcctcatg aatgtctttt cactacccat ttgcttatcc tgcatctctc agccttgact
     3961 ccactcagtt ctcttgctta gagataccac ctttcccctg aagtgttcct tccatgtttt
     4021 acggcgagat ggtttctcct cgcctggcca ctcagcctta gttgtctctg ttgtcttata
     4081 gaggtctact tgaagaagga aaaacagggg gcatggtttg actgtcctgt gagcccttct
     4141 tccctgcctc ccccactcac agtgacccgg aatccctcga catggcagtc tagagcatgg
     4201 ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
     4261 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4321 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctggcgtaa
     4381 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4441 gcgattccgt tgcaatggct ggcggtaata ttgttctgga tattaccagc aaggccgata
     4501 gtttgagttc ttctactcag gcaagtgatg ttattactaa tcaaagaagt attgcgacaa
     4561 cggttaattt gcgtgatgga cagactcttt tactcggtgg cctcactgat tataaaaaca
     4621 cttctcagga ttctggcgta ccgttcctgt ctaaaatccc tttaatcggc ctcctgttta
     4681 gctcccgctc tgattctaac gaggaaagca cgttatacgt gctcgtcaaa gcaaccatag
     4741 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4801 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4861 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4921 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4981 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5041 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5101 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5161 aacgcgaatt ttaacaaaat attaacgctt acaatttaaa tatttgctta tacaatcttc
     5221 ctgtttttgg ggcttttctg attatcaacc ggggtacata tgattgacat gctagtttta
     5281 cgattaccgt tcatcgattc tcttgtttgc tccagactct caggcaatga cctgatagcc
     5341 tttgtagaga cctctcaaaa atagctaccc tctccggcat gaatttatca gctagaacgg
     5401 ttgaatatca tattgatggt gatttgactg tctccggcct ttctcacccg tttgaatctt
     5461 tacctacaca ttactcaggc attgcattta aaatatatga gggttctaaa aatttttatc
     5521 cttgcgttga aataaaggct tctcccgcaa aagtattaca gggtcataat gtttttggta
     5581 caaccgattt agctttatgc tctgaggctt tattgcttaa ttttgctaat tctttgcctt
     5641 gcctgtatga tttattggat gttggaatcg cctgatgcgg tattttctcc ttacgcatct
     5701 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     5761 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5821 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5881 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     5941 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6001 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6061 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6121 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6181 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6241 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6301 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6361 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6421 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6481 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6541 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6601 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6661 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6721 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6781 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6841 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6901 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6961 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7021 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7081 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7141 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7201 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7261 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7321 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7381 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7441 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7501 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7561 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7621 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7681 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7741 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7801 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7861 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7921 caaaccgcct ctccccgcgc gttggccgat tcatta
//
