LOCUS       sequence-347130-        5969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  neural activity dependent release of sbGLuc from presynaptic 
            vesicles.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5969)
  AUTHORS   Prakash M, Murphy J, St Laurent R, Friedman N, Crespo EL, Bjorefeldt
            A, Pal A, Bhagat Y, Kauer JA, Shaner NC, Lipscombe D, Moore CI, 
            Hochgeschwender U
  TITLE     Selective control of synaptically-connected circuit elements by 
            all-optical synapses.
  JOURNAL   Commun Biol. 2022 Jan 11;5(1):33. doi: 10.1038/s42003-021-02981-7.
  PUBMED    35017641
REFERENCE   2  (bases 1 to 5969)
  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: "Commun 
            Biol."; date: "2022-01-11"; volume: "5(1):33. doi"; pages: " 
            10.1038/s42003-021-02981-7"
COMMENT     Sequence Label: pAAV-hSyn-hPOMC1-26-sbGLuc-P2A-dTomato
FEATURES             Location/Qualifiers
     source          1..5969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   107..247
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   107..236
                     /label=AAV2 ITR
     rep_origin      322..777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(409..428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     619..640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(794..813)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     913..935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(973..991)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1059..1163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1164..1232,1233..2024)
                     /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(1382..1401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2195..2783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2684..2703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2845..2974
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3019..3466
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             4088..4144
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4163..4864
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(4296..4314)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4564..4583
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4758..4781
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             4892..4909
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4940..5528
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4993..5013)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5411..5422)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(5560..5969,1..67)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5759..5778)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
       61 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
      121 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
      181 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
      241 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
      301 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      361 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
      421 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      481 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
      541 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      601 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
      661 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
      721 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
      781 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
      841 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
      901 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
      961 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     1021 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1081 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1141 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1201 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1261 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1321 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1381 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1441 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1501 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     1561 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     1621 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     1681 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     1741 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     1801 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     1861 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     1921 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     1981 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2041 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2101 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2161 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2221 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2281 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2341 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2401 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2461 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2521 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2581 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2641 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2701 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2761 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2821 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
     2881 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     2941 gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgtgt gtctagactg
     3001 cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg
     3061 tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat
     3121 tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact
     3181 gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg
     3241 cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg
     3301 ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga
     3361 gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca
     3421 gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga gaaggtaccg
     3481 gatccccacc atgccgagat cgtgctgcag ccgctcgggg gccctgttgc tggccttgct
     3541 gcttcaggcc tccatggaag tgcgtggcaa gcccaccgag aacaacgaag acttcaacat
     3601 cgtggccgtg gccagcaact tcgcgaccac ggatctcgat gctgaccgcg ggaagttgcc
     3661 cggcaagaag ctgccgctgg aggtgctcaa agagctggaa gccaatgccc ggaaagctgg
     3721 ctgcaccagg ggctgtctga tctgcctgtc ccacatcaag tgcacgccca agatgaagaa
     3781 gttcatccca ggacgctgcc acacctacga aggcgacaaa gagtccgcac agggcggcat
     3841 aggcgaggcg atcgtcgaca ttcctgagat tcctgggttc aaggacttgg agcccctgga
     3901 gcagttcatc gcacaggtcg atctgtgtgt ggactgcaca actggctgcc tcaaagggct
     3961 tgccaacgtg cagtgttctg acctgctcaa gaagtggctg ccgcaacgct gtgcgacctt
     4021 tgccagcaag atccagggcc aggtggacaa gatcaagggg gccggtggtg acctcgaggg
     4081 aagcggagcc actaacttct ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg
     4141 gccagcggcc gcacccgggc caatggtgag caagggcgag gaggtcatca aagagttcat
     4201 gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     4261 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     4321 ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta
     4381 cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa
     4441 gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc
     4501 cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga
     4561 cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     4621 ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca
     4681 ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta
     4741 ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     4801 acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta
     4861 caagtaacat atgctctaga caattgccgg tcatcatcac catcaccatt gagtttaatt
     4921 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     4981 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5041 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5101 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5161 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5221 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5281 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     5341 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
     5401 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5461 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     5521 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     5581 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     5641 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     5701 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     5761 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     5821 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     5881 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     5941 ctggtctcca actcctaatc tcaggtgat
//
