LOCUS       sequence-91188-e        6685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of Mac-GFP under the Syn promoter, in 
            floxed/reversed (Cre-dependent) manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6685)
  AUTHORS   Chow BY, Han X, Dobry AS, Qian X, Chuong AS, Li M, Henninger MA, 
            Belfort GM, Lin Y, Monahan PE, Boyden ES
  TITLE     High-performance genetically targetable optical neural silencing by 
            light-driven proton pumps.
  JOURNAL   Nature. 2010 Jan 7. 463(7277):98-102.
  PUBMED    20054397
REFERENCE   2  (bases 1 to 6685)
  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: "Nature. 
            2010 Jan 7. 463(7277):98-102."
COMMENT     Sequence Label: pAAV-Syn-FLEX-Mac-GFP
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        355..802
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    818..851
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    914..947
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(961..1674,1675..1677))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(964..1674,1675..1677))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1006..1027)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1355..1374
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1614..1635
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2648..2681)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2752..2785)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2818..3406
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2871..2891)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3289..3300)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3426..3902
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3625..3644)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3948..4088
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3948..4077
                     /label=AAV2 ITR
     rep_origin      4163..4618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4250..4269)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4460..4481
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4635..4654)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4754..4776
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4814..4832)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4900..5004
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5005..5073,5074..5865)
                     /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(5223..5242)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6036..6624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6525..6544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttgcaa agatggataa agttttaaac
      181 agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc
      241 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
      301 ggtcggcagc aaatggttaa ttaagtgtct agactgcaga gggccctgcg tatgagtgca
      361 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      421 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      481 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      541 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      601 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      661 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      721 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      781 cgtgtcgtgc ctgagagcgc agtcgagaag aattcccata acttcgtata aagtatccta
      841 tacgaagtta tatcaaaata ggaagaccaa tgcttcacca tcgacccgaa ttgccaagca
      901 tcaccatcga cccataactt cgtataatgt atgctatacg aagttatact agctagctca
      961 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     1021 gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta
     1081 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     1141 ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc
     1201 gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt
     1261 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1321 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1381 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1441 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1501 ggtgggccag ggcacgggca gcttgccggt ggtgcaaatg aacttcaggg tcagcttgcc
     1561 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1621 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcactgc
     1681 tactaccggt cgggcgccgt cgtcctcgcc gatcctgatg gcgccctccc tgttcaggcc
     1741 gttggcccag aagccgttca gctccacgtc gctctccctc aggttggcgt gggtcaccag
     1801 cagccaagct ccgaacactc ccttagccag cacgtccagc acagcgtaag cgatgatctc
     1861 tccgtccact ccgatcttcc tagctccgtc agccagtccc cacacgattg ggtaagcggt
     1921 ccacaggatc agggtgtaag ctccgatggc cacgaagaag ctcctcagct tctcgccctt
     1981 caccctggcg ttggcgccgc cgttcagcac caggtgccac accacgaaga tgtaggcgat
     2041 gcaggcgatg gtgtaccatc cccacttctg tggggttccc tcgctgccga aagcagcgaa
     2101 caggccggtc agcaccatga tcaggtcagc cacgatggcc atgaagatgt gggctccgct
     2161 cattccagcc agcagtccca ggtccagcag cagcagtggg gtggtgatag cccagtcgat
     2221 gtacctggcg tagtacacct gtcggtacac ggtctcgtag gtgtcgggca cgtggtcgtg
     2281 ctgggtcctg atcacgatct tgttcagggc cactccgtgg ccggtagcca tagcgaagta
     2341 gctcagggcg gcggtcaggg tgatgatggt ggtgatcacg tggtacagcc tcctgttcac
     2401 ggggatcttc cagctcaggg cggtgaaggc ggcgctggcg atcagcatca gcacgaacac
     2461 cacccacagg gtcttgctgc cgctgtcgcc cacggtctcg tagatggggg tgtcgggcag
     2521 cacggttggc acgggggcca cgtgggttgg ctgggcgctc tgggtagcgg ttggcagtgg
     2581 gaacagctgg ctggtcttca tcagcacctc ctcgaactgg tccacgatca tggtggcggt
     2641 acctagtata acttcgtata ggatacttta tacgaagtta tcattgggat tcttcctatt
     2701 ttgatccaag catcaccatc gaccctctag tccagatctc accatcgacc cataacttcg
     2761 tatagcatac attatacgaa gttatgtccc tcgaagaggt tcgcgaagct tatcgataat
     2821 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2881 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2941 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3001 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3061 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3121 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3181 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3241 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3301 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3361 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccggctc
     3421 gagacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     3481 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     3541 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     3601 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     3661 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     3721 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     3781 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     3841 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     3901 ttctcgagct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     3961 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4021 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4081 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4141 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4201 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4261 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4321 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4381 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4441 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4501 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4561 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4621 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4681 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4741 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4801 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4861 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4921 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4981 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5041 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5101 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5161 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5221 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5281 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5341 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5401 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5461 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5521 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5581 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5641 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5701 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5761 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5821 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5881 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5941 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6001 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6061 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6121 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6181 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6241 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6301 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6361 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6421 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6481 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6541 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6601 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6661 ttttgctggc cttttgctca catgt
//
