LOCUS       sequence-317905-        6691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate Adeno-Associated viruses for expression of psychLight2 
            under synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6691)
  AUTHORS   Dong C, Ly C, Dunlap LE, Vargas MV, Sun J, Hwang IW, Azinfar A, Oh 
            WC, Wetsel WC, Olson DE, Tian L
  TITLE     Psychedelic-inspired drug discovery using an engineered biosensor.
  JOURNAL   Cell. 2021 Apr 8. pii: S0092-8674(21)00374-3. doi: 
            10.1016/j.cell.2021.03.043.
  PUBMED    33915107
REFERENCE   2  (bases 1 to 6691)
  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: "Cell. 2021 
            Apr 8. pii: S0092-8674(21)00374-3. doi: 10.1016/j.cell.2021.03.043."
COMMENT     Sequence Label: pAAV_hSynapsin_psychLight2
FEATURES             Location/Qualifiers
     source          1..6691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     promoter        470..917
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     2040..2061
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2167..2188)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2428..2447)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3079..3667
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3132..3152)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3550..3561)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3670..3686)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3671..3687)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3716..3837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3727..3746)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3781..3800
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3999..4128
                     /label=AAV2 ITR
     primer_bind     complement(4147..4164)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4147..4163)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4156..4178)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4305..4760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4392..4411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4602..4623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4786..4890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4891..4959,4960..5751)
                     /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(5109..5128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5922..6510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6411..6430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg
      481 gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac
      541 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      601 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      661 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      721 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      781 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      841 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
      901 cgtgcctgag agcgcagtcg aattcaagct gctagcaagg atccacccgc caccatgaag
      961 acgatcatcg ccctgagcta catcttctgc ctggtgttcg ccgactacaa ggacgatgat
     1021 gacgccatgg acatactttg tgaagagaat acttcactct cttctactac taactctctt
     1081 atgcaactga acgatgatac ccgattgtac tcaaacgact tcaattccgg cgaagcgaac
     1141 accagtgacg cattcaactg gactgtcgat tctgaaaaca gaactaatct gtcatgcgag
     1201 ggttgtctta gtccctcttg tctcagcctg ttgcacctcc aggaaaagaa ctggtcagca
     1261 ctgctcactg cggtagtgat aatactcact attgctggca atattctcgt aattatggca
     1321 gtctccttgg agaagaaact ccaaaacgcc acaaattatt ttcttatgag ccttgccatc
     1381 gcagatatgc tcttgggatt tttggtgatg cctgtgagta tgctcacgat actgtatgga
     1441 tatcgctggc ctctgccgtc taaactttgc gctgtgtgga tttacttgga tgtccttttt
     1501 tcaactgcga gtattatgca tctttgcgcc attagtcttg ataggtatgt ggctatccaa
     1561 aatcctaagc accattcccg ctttaatagt agaactaagg cttttctgaa aataatagca
     1621 gtgtggacca tatctgtcgg cataagcatg cctatccccg tatttggact tcaagatgac
     1681 tcaaaggtat tcaaagaagg gtcatgtctg ctggccgatg acaatttcgt gcttattggc
     1741 agcttcgtca gtttcttcat tcctttgaca atcatggtga ttacctactt tcttacgatt
     1801 aaatctttgc aaaagcagct gagctcaggt tataacgtct atatcaaggc cgacaagcag
     1861 aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     1921 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1981 aaccactacc tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac
     2041 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2101 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg
     2161 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2221 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2281 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2341 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2401 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2461 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2521 gacggcaaca tcctggggca caagctggag tacaacttta atatgcatga ccaactgaat
     2581 gagcaaaagg cttgtaaggt actcggcata gtcttctttc tgtttgtggt gatgtggtgt
     2641 cccttcttta taacgaatat catggcagtg atctgcaagg aatcatgcaa tgaggatgtg
     2701 atcggggcac ttctgaacgt tttcgtgtgg atagggtatc tgtcaagtgc tgtgaaccca
     2761 ctggtctata ccttgtttaa taagacatac cgctcagcct tttcacggta tattcaatgt
     2821 cagtataagg aaaacaagaa acctctgcaa cttattcttg tgaacactat ccctgccctg
     2881 gcttataagt catcacagtt gcagatgggc cagaaaaaaa attccaagca ggacgcgaag
     2941 acaacagaca acgattgtag tatggttgcc ctcggcaagc agcacagtga agaagcgagc
     3001 aaagacaata gtgatggcgt aaacgaaaaa gtcagttgtg tattctgcta cgagaacgag
     3061 gtgtaaaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3121 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3181 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3241 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3301 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3361 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3421 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3481 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3541 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3601 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3661 tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga
     3721 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3781 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3841 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3901 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     3961 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4021 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4081 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc
     4141 taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4201 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4261 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4321 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4381 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4441 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4501 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4561 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4621 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4681 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4741 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4801 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4861 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4921 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4981 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5041 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5101 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5161 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5221 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5281 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5341 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5401 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5461 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5521 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5581 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5641 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5701 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5761 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5821 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5881 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5941 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6001 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6061 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6121 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6181 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6241 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6301 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6361 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6421 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6481 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6541 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6601 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6661 cgcagcgagt cagtgagcga ggaagcggaa g
//
