LOCUS       sequence-408011-        6312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate Adeno-Associated viruses for expression of dLight1.2 
            under synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6312)
  AUTHORS   Patriarchi T, Cho JR, Merten K, Howe MW, Marley A, Xiong WH, Folk 
            RW, Broussard GJ, Liang R, Jang MJ, Zhong H, Dombeck D, von Zastrow 
            M, Nimmerjahn A, Gradinaru V, Williams JT, Tian L
  TITLE     Ultrafast neuronal imaging of dopamine dynamics with designed 
            genetically encoded sensors.
  JOURNAL   Science. 2018 Jun 29;360(6396). pii: science.aat4422. doi: 
            10.1126/science.aat4422. Epub 2018 May 31.
  PUBMED    29853555
REFERENCE   2  (bases 1 to 6312)
  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: "Science."; 
            date: "2018-06-29"; volume: "360(6396). pii: science.aat4422. doi"; 
            pages: " 10.1126/science.aat4422. Epub 2018 May 31"
COMMENT     Sequence Label: pAAV-hSyn-dLight1.2
FEATURES             Location/Qualifiers
     source          1..6312
                     /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)"
     primer_bind     1657..1678
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1784..1805)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2045..2064)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2750..3338
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2803..2823)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3221..3232)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3341..3357)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3342..3358)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3387..3508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3398..3417)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3452..3471
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3670..3799
                     /label=AAV2 ITR
     rep_origin      complement(4036..4624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4116..4135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4795..5586,5587..5655))
                     /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     5418..5437
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5656..5760)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5786..6241)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5923..5944)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6135..6154
                     /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 tcagcaccgc 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 cgaattcaag ctgctagcaa ggatcctgcg cgccatgaag acgatcatcg ccctgagcta
      721 catcttctgc ctggtgttcg ccgactacaa ggacgatgat gacgccatga ggactctgaa
      781 cacctctgcc atggacggga ctgggctggt ggtggagagg gacttctctg ttcgtatcct
      841 cactgcctgt ttcctgtcgc tgctcatcct gtccacgctc ctggggaaca cgctggtctg
      901 tgctgccgtt atcaggttcc gacacctgcg gtccaaggtg accaacttct ttgtcatctc
      961 cttggctgtg tcagatctct tggtggccgt cctggtcatg ccctggaagg cagtggctga
     1021 gattgctggc ttctggccct ttgggtcctt ctgtaacatc tgggtggcct ttgacatcat
     1081 gtgctccact gcatccatcc tcaacctctg tgtgatcagc gtggacaggt attgggctat
     1141 ctccagccct gcccggtatg agagaaagat gacccccaag gcagccttca tcctgatcag
     1201 tgtggcatgg accttgtctg tactcatctc cttcatccca gtgcagctca gctggcacaa
     1261 ggcaaaaccc acaagcccct ctgatggaaa tgccacttcc ctggctgaga ccatagacaa
     1321 ctgtgactcc agcctcagca ggacatatgc catctcatcc tctgtaatca gcttttacat
     1381 ccctgtggcc atcatgattg tcacctacac caggatctac aggattgctc agaaactgag
     1441 ctcactcatt aacgtctata tcaaggccga caagcagaag aacggcatca aggcgaactt
     1501 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa
     1561 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc
     1621 caaactttcg aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1681 cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat
     1741 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1801 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg
     1861 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1921 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1981 gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt
     2041 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2101 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2161 gctggagtac aacaatcatg accaactgaa aagagaaact aaagtcctga agactctgtc
     2221 ggtgatcatg ggtgtgtttg tgtgctgttg gctacctttc ttcatcttga actgcatttt
     2281 gcccttctgt gggtctgggg agacgcagcc cttctgcatt gattccaaca cctttgacgt
     2341 gtttgtgtgg tttgggtggg ctaattcatc cttgaacccc atcatttatg cctttaatgc
     2401 tgattttcgg aaggcatttt caaccctctt aggatgctac agactttgcc ctgcgacgaa
     2461 taatgccata gagacggtga gtatcaataa caatggggcc gcgatgtttt ccagccatca
     2521 tgagccacga ggctccatct ccaaggagtg caatctggtt tacctgatcc cacatgctgt
     2581 gggctcctct gaggacctga aaaaggagga ggcagctggc atcgccagac ccttggagaa
     2641 gctgtcccca gccctatcgg tcatattgga ctatgacact gacgtctcta tggagaagat
     2701 ccaacccatc acacaaaacg gtcagcaccc aacctgaaag cttatcgata atcaacctct
     2761 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2821 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2881 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2941 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3001 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3061 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3121 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     3181 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3241 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3301 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc
     3361 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     3421 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3481 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3541 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     3601 aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca
     3661 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3721 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     3781 tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3841 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3901 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3961 taacgcagga aagaacatgt gagcaaaagg ccagcaaaaa gccaggaacc gtaaaaaggc
     4021 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4081 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4141 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4201 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4261 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4321 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4381 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4441 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4501 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4561 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4621 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4681 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4741 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4801 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4861 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4921 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4981 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5041 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5101 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5161 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5221 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5281 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5341 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5401 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5461 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5521 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5581 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5641 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5701 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5761 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     5821 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5881 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5941 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     6001 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     6061 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     6121 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     6181 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     6241 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6301 ctattacgcc ag
//
