LOCUS       sequence-169763-        6078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of Chronos-GFP under the Syn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6078)
  AUTHORS   Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK,
            Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, 
            Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, 
            Constantine-Paton M, Wong GK, Boyden ES
  TITLE     Independent optical excitation of distinct neural populations.
  JOURNAL   Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 
            2014 Feb 9.
  PUBMED    24509633
REFERENCE   2  (bases 1 to 6078)
  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"; doi: 
            "10.1038/nmeth.2836"; journalName: "Nat Methods"; date: "2014-03"; 
            volume: "11"; issue: "3"; pages: "338-46"
COMMENT     Sequence Label: pAAV-Syn-Chronos-GFP
FEATURES             Location/Qualifiers
     source          1..6078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      8..17
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(30..477)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   577..706
                     /label=AAV2 ITR
     rep_origin      778..1233
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(865..884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1075..1096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1259..1363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1364..1432,1433..2224)
                     /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(1582..1601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2395..2983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2884..2903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3220..3349
                     /label=AAV2 ITR
     polyA_signal    3519..3726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3767..4355
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3873..3884
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4282..4302
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4380..5093,5094..5096))
                     /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(4383..5093,5094..5096))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4425..4446)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4774..4793
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5033..5054
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 gctgtttcca tggtggcgga tcctctcgac tgcgctctca ggcacgacac gactcctccg
       61 ctgcccaccg cagactgagg cagcgctgag tcgccggcgc cgcagcgcag atggtcgcgc
      121 ccgtgccccc ctatctcgcg cctcgcgtgg tgcggtccgg ctgggccggc ggcggcgcgg
      181 acgcgaccaa ggtggccggg aaggggagtt tgcgggggac cggcgagtga cgtcagcgcg
      241 ccttcagtgc tgaggcggcg gtggcgcgcg ccgccaggcg ggggcgaagg cactgtccgc
      301 ggtgctgaag ctggcagtgc gcacgcgcct cgccgcatcc tgtttcccct ccccctctct
      361 gataggggat gcgcaatttg gggaatgggg gttgggtgct tgtccagtgg gtcggggtcg
      421 gtcgtcaggt aggcaccccc accccgcctc atcctggtcc taaaacccac ttgcactcat
      481 acgcagggcc ctctgcagtc tagattaatt aacctacgat cttcctagag catggctacg
      541 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
      601 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
      661 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgg cgtaatagcg
      721 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
      781 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      841 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      901 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      961 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1021 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1081 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1141 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1201 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     1261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1321 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1381 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1441 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1501 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1561 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     1621 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1681 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1741 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1801 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1861 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     1921 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1981 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2041 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2101 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2161 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2221 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2521 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3061 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3121 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3181 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tgcgcgctcg ctcgctcact
     3241 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     3301 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg
     3361 attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcct tatcgatttt
     3421 accacatttg tagaggtttt acttgcttta aaaaacctcc cacatctccc cctgaacctg
     3481 aaacataaaa tgaatgcaat tgtacagtcc ggaccgcatc cccagcatgc ctgctattgt
     3541 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
     3601 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
     3661 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
     3721 gcacagtcga gctttatgag cggccgcccg ggtcgacggt atcgatgcgg ggaggcggcc
     3781 caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc
     3841 ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag
     3901 gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccaaggaaa ggacgatgat
     3961 ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
     4021 gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg
     4081 gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca
     4141 gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac
     4201 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     4261 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     4321 aatctttcac aaattttgta atccagaggt tgattatcga taagcttgat atcgaattct
     4381 tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     4441 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag
     4501 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     4561 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     4621 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     4681 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     4741 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     4801 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     4861 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     4921 gtgggccagg gcacgggcag cttgccggtg gtgcaaatga acttcagggt cagcttgccg
     4981 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     5041 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcactgct
     5101 actaccggtg ccgccactcc tccctcctcc tcctcgtcga cgaaggtctc gatctccatg
     5161 ttctcgccgg ccacgttgat ggtggttgtc ttccggatgt cgccgtggat caggatgtgc
     5221 tcgtggatct tcactctcag gtggtggccc aggaagcccc aagtgttctt gctgatcaga
     5281 tccaggatca ggtggccgat tccgctggtg taaggtgtga tcaggcccag tccctcgtgt
     5341 ccggcgatga acatcacggg gaacatcagc caagagcaga agaagacgta ggccatgatc
     5401 ttgcaaatct tccggcagac gcctttgggc agggtgtggt agctctcgat atacaccttg
     5461 gcgatctgga agaagcaagt cacgccgtag aacaggccga tcatgaagaa caggatcttc
     5521 agggggccct ttgtaaaggc ggctgtgatc ccccacacga tgttgccgat gtcggtcacc
     5581 aggatggtca tggtccgctt gctgtactct tcgtgcagtc cggtcaggtt gctcaggtgg
     5641 atcaggatca cggggcaagt caggagccac atgctgtacc gcagccaaat cacggctcct
     5701 ccgttggtct ggtacactgt ggctgggctg tcgacctcgt ggaacagctc gatgaagcac
     5761 ttgacgagct cgatcacgca gacgtagacc tcctcccagc cggttgtggc cttgaagctg
     5821 tgccaaccgt agaagatgag ctgcacgata gccacgatca cggtgaacca ctggagtccc
     5881 actgcgatct tgtgctgcag ctcggttccg tggttgatgt gggggaagca accgtgatcg
     5941 gctccggctg ttgtggcgtt agaggtttct ccgtgagcgt ctgctgctgg tgtcaccact
     6001 gctgcggcgc tcagcaggcc tctaattgtg gcttcggcgc taggcacggc tgagataaag
     6061 gcgtgggtca ttgtggcg
//
