LOCUS       sequence-244599-        5918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV based broad expression of eTsChR in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5918)
  AUTHORS   Farhi SL, Parot VJ, Grama A, Yamagata M, Abdelfattah AS, Adam Y, Lou
            S, Jun Kim J, Campbell RE, Cox DD, Cohen AE
  TITLE     Wide-area all-optical neurophysiology in acute brain slices.
  JOURNAL   J Neurosci. 2019 Apr 5. pii: JNEUROSCI.0168-19.2019. doi: 
            10.1523/JNEUROSCI.0168-19.2019.
  PUBMED    30952812
REFERENCE   2  (bases 1 to 5918)
  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: "J Neurosci.
            2019 Apr 5. pii: JNEUROSCI.0168-19.2019. doi: 
            10.1523/JNEUROSCI.0168-19.2019."
COMMENT     Sequence Label: SLF045: AAV-hSyn-eTsChR-GFP
FEATURES             Location/Qualifiers
     source          1..5918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        212..659
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1627..1629,1630..2343)
                     /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             join(1627..1629,1630..2340)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1669..1690)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1930..1949)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2277..2298
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2356..2944
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2409..2429)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2827..2838)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2947..2963)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2948..2964)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2993..3114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3004..3023)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3058..3077
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3276..3405
                     /label=AAV2 ITR
     rep_origin      complement(3642..4230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3722..3741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4401..5192,5193..5261))
                     /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     5024..5043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5262..5366)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5392..5847)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5529..5550)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5741..5760
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg accaggatga
      241 ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc
      301 ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag
      361 gcgcgtgcgc actgccagct tcaacaccgc ggacagtgcc ttcgcccccg cctggcggcg
      421 cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa
      481 ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc
      541 acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact
      601 cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt
      661 cgaattcggg gatccaccat gttcgccatc aaccccgaat atatgaatga gactgtgctg
      721 ctggacgaat gcaccccaat ctacctggat attggaccac tgtgggagca ggtggtggct
      781 agggtgactc agtggttcgg agtgatcctg tctctggtgt ttctgatcta ctacatctgg
      841 aacacttaca aggctacctg tgggtgggag gaactgtacg tgtgcacagt ggagttctgt
      901 aagatcatca tcgaactgta cttcgagtat actccccctg ccatgatctt ccagacaaat
      961 ggacaagtga ctccctggct gcggtatgct gagtggctgc tgacatgccc tgtgatcctg
     1021 attcacctgt ctaacattac tgggctgaat gacgattaca gtggccgcac catgagcctg
     1081 atcacatccg acctgggcgg aatttgtatg gctgtgaccg ccgctctgag caagggatgg
     1141 ctgaaagccc tgttctttgt gatcgggtgc ggctacggag ccagcacctt ctacaacgcc
     1201 gcttgtatct acattgagtc ctactatacc atgccacagg gaatttgcag gagactggtg
     1261 ctgtggatgg ccggggtgtt ctttacatcc tggttcatgt ttcctggcct gttcctggct
     1321 ggaccagaag gaacccaggc cctgtcttgg gctggaacca caatcggaca tacagtggcc
     1381 gacctgctga gtaagaacgc ttggggaatg atcgggcact ttctgagggt ggagatccac
     1441 aagcatatca tcatccatgg agatgtgcgg cgccccgtga cagtgaaggc tctggggagg
     1501 caggtgagcg tgaattgttt cgtggacaaa gaggaagagg aagaggatga gagaatcgcg
     1561 gccgccaaga gcaggatcac cagcgagggc gagtacatcc ccctggacca gatcgacatc
     1621 aacgtggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1681 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1741 tacggcaagc tgaccctgaa gttcatttgc accaccggca agctgcccgt gccctggccc
     1801 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1861 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1921 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1981 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2041 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2101 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2161 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2221 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2281 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2341 tagaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2401 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2461 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2521 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2581 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2641 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2701 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2761 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2821 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2881 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2941 ccgcatcgat accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac
     3001 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3061 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3121 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     3181 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     3241 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     3301 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3361 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     3421 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3481 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3541 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3601 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3661 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3721 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3781 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3841 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3901 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3961 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4021 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4081 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4141 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4681 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     5401 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5461 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5521 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5581 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5641 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5701 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5761 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5821 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     5881 aagggcgatc ggtgcgggcc tcttcgctat tacgccag
//
