LOCUS       sequence-393500-        7218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for chemogenetically silencing and visualizing 
            subpopulations of neurons in the retina.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7218)
  TITLE     Intrsectional chemogenetic viruses for retinal neurobiology
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7218)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-hSyn-con/fon DREADD Gi-mCherry
FEATURES             Location/Qualifiers
     source          1..7218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    636..669
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     complement(990..1009)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1272..1305)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1306..1339)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     regulatory      2000..2009
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2151..2169)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    2251..2284
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2285..2318
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     regulatory      3231..3240
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3253..3286)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    3345..3933
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3398..3418)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3816..3827)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3965..4441
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4164..4183)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4481..4621
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4481..4610
                     /label=AAV2 ITR
     rep_origin      4696..5151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4783..4802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4993..5014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5168..5187)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5287..5309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5347..5365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5433..5537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5538..5606,5607..6398)
                     /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(5756..5775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6569..7157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7058..7077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccgaagt tcctattctt caaatagtat
      661 aggaacttcg aagttcctat tcttcaaaag gtataggaac ttcggcgcgc cttacttgta
      721 cagctcgtcc atgccgccgg tggagtggcg gccctcggcg cgttcgtact gttccacgat
      781 ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt aggcgccggg
      841 cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc
      901 gtccttcagc ttcagcctct gcttgatctc gcccttcagg gcgccgtcct cggggtacat
      961 ccgctcggag gaggcctccc agcccatggt cttcttctgc attacggggc cgtcggaggg
     1021 gaagttggtg ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca gggaggagtc
     1081 ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc acttgaagcc
     1141 ctcggggaag gacagcttca agtagtcggg gatgtcggcg gggtgcttca cgtaggcctg
     1201 tgggacgaca tgacttaacc agttaaccat gttctctgaa catgagaagg gctggccact
     1261 ctccagtcga cataacttcg tataggatac tttatacgaa gttatataac ttcgtatagt
     1321 ataccttata cgaagttata ctagttgtgc cccccctttt ttttataaaa ttgtattaat
     1381 gttatataca tatctcctgt atgtgaccca tgtgcttatg actctatttc tcatgtgttt
     1441 aggcagtgcc acccagaaca ccaaggaacg cccagccaca gagctgtcca ccacagaggc
     1501 caccacgccc gccatgcccg cccctcccct gcagccgcgg gccctcaacc cagcctccag
     1561 atggtccaag atccagattg tgacgaagca gacaggcaat gagtgtgtga cagccattga
     1621 gattgtgcct gccacgccgg ctggcatgcg ccctgcggcc aacgtggccc gcaagttcgc
     1681 cagcatcgct cgcaaccagg tgcgcaagaa gcggcagatg gcggcccggg agcgcaaagt
     1741 gacacgaacg atctttgcca ttctgctggc cttcatcctc acctggacgc cctacaacgt
     1801 catggtcctg gtgaacacct tctgccagag ctgcatccct gacacggtgt ggtccattgg
     1861 ctactggctc tgctacgtca acagcaccat caaccctgcc tgctatgctc tgtgcaacgc
     1921 cacctttaaa aagaccttcc ggcacctgct gctgtgccag tatcggaaca tcggcactgc
     1981 caggcgggat ccaccggtcg ccaccatggt gagcaagggc gaggaggata acatggccat
     2041 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     2101 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     2161 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     2221 cggctccaag gtgagtacag gaggggatcc ataacttcgt ataaagtatc ctatacgaag
     2281 ttatataact tcgtataagg tatactatac gaagttatgc tagcaacccc gacacttacc
     2341 tgagctggac tcattggaag tgtccttatc agccacgggg cgcggtggcg gtggcagcgc
     2401 tggcgggggg gcctcctcca gcttgccatt gcgcagctcc tcccgggcgg cctccccggg
     2461 cgggggcttc ttgacgctct gcttcattag tgggctcttg aggaaggcca gcgtcttggc
     2521 tttcttctcc ttcgggccct cgggccggtg cttgtggact cggctgcgac tggccaggga
     2581 gatgtggatg tacagcaccg tcatgatgac cacaggcagg tagaagccag caatggctgt
     2641 gccaaaggtc actgctgggt tggacaggaa ctggatgaag cactggttgt cgggcaccgt
     2701 ccgcttaccc accacaaact gccagaacaa gatggcaggc gcccagagca cgaaggacag
     2761 tacccaggca gcagcaatca tgaggcctgc catcttggtg gtgcgccggg cagggtaggt
     2821 gagaggcttg gtgacgcaga agtagcggtc aaagctgatg atgagaaggt tcatgacgga
     2881 ggcgttgctc accacgcagt ccagggccag ccacaggtcg cagaccacgg cgcccagggg
     2941 ccagtagccc ttgatgatgt acacggtgta gaggttcatg gagaaggcgc ctatgatgag
     3001 atcagcacac gccaggctga agaggaagta gttgttgact gtctgcagct gcctgttgac
     3061 cttgatggac agcatcacca ggatgttgcc cacgacagtc accaggctca gggagcctgt
     3121 cactgtggca atgaagacca tttccaccgt ctcatagcga ttgtgggatg atgacgtgac
     3181 caggcgcacg gactgattgc ccgagctgcc attgacaggt gtgaagttgg ccatggtggc
     3241 ggtggcgagc tcgaagttcc tatactattt gaagaatagg aacttcgaag ttcctatacc
     3301 ttttgaagaa taggaacttc gaattcgata tcaagcttat cgataatcaa cctctggatt
     3361 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3421 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3481 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3541 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3601 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3661 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3721 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     3781 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3841 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3901 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     3961 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     4021 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     4081 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     4141 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     4201 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     4261 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     4321 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     4381 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     4441 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     4501 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4561 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4621 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4681 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4741 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     4801 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4861 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4921 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4981 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     5041 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     5101 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     5161 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5221 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5281 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     5341 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5401 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5461 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5521 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5581 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5641 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5701 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5761 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5821 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5881 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5941 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6001 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6061 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6121 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6181 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6241 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6301 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6361 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6421 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6481 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6541 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6601 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6661 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6721 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6781 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6841 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6901 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6961 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7021 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7081 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7141 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7201 ggccttttgc tcacatgt
//
