LOCUS       sequence-204352-        6446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a soma-targeted GtACR2 in Cre-expressing neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6446)
  AUTHORS   Mahn M, Gibor L, Patil P, Cohen-Kashi Malina K, Oring S, Printz Y, 
            Levy R, Lampl I, Yizhar O
  TITLE     High-efficiency optogenetic silencing with soma-targeted 
            anion-conducting channelrhodopsins.
  JOURNAL   Nat Commun. 2018 Oct 8;9(1):4125. doi: 10.1038/s41467-018-06511-8.
  PUBMED    30297821
REFERENCE   2  (bases 1 to 6446)
  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: "Nat 
            Commun."; date: "2018-10-8"; volume: "9(1):4125. doi"; pages: " 
            10.1038/s41467-018-06511-8"
COMMENT     Sequence Label: pAAV_hSyn1-SIO-stGtACR2-FusionRed
FEATURES             Location/Qualifiers
     source          1..6446
                     /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    complement(636..669)
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(877..1572)
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     regulatory      2511..2520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2527..2560
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2573..3161
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2626..2646)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3044..3055)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3193..3669
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3392..3411)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3709..3849
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3709..3838
                     /label=AAV2 ITR
     rep_origin      3924..4379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4011..4030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4221..4242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4396..4415)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4515..4537
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4575..4593)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4661..4765
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4766..4834,4835..5626)
                     /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(4984..5003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5797..6385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6286..6305
                     /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 gtacctaccg ttcgtatagc atacattata
      661 cgaagttatg aattcttact taaaccgcgt agcctctgga aaatcggtgg cgcatgagat
      721 aaaagagtca atagaactca tggacctcat gtcgatcacg ccctctgttc ttgcaggaag
      781 aggtgccaca ggggagggca ttgaggacat ttccagctcc tcagggggtt tactctgggg
      841 ggccatctct ttggtgttca gaataggctg ggactgttta cctccatcac cagcgccacc
      901 agtagagtat ctggccacag ccacctcgtg ctgctcgacg tatgtctcat cgtcggcctc
      961 cttgattctt tccagtctgt ggtccacgtt gtagacgccg ggcatcttga ggttcgtagc
     1021 gggtttcttg gatctgtatg ttgtctcaag gttgcagatc aggtgtcctc ctcccacgag
     1081 cttcagagcc atgtcacatg cgccttccag tcctccgtca gcagggtaca tcgtctcggt
     1141 ggaggcctcc cagccgagtg ttttcttctg catcacaggg ccgttggctg ggaagttcac
     1201 ccctctaacc ttgacgttgt agatgaggca gccgtcctgg aggctggtgt cctgggtagc
     1261 ggtcagcacg cccccgtctt cgtatgtggt gactctctcc catgtgaagc cctcagggaa
     1321 ggactgctta aagaagtcgg ggatgcccgg agggtgcttg atgaaggttc tgctgccgta
     1381 catgaagctg gtagccagga tgtcgaaggc gaaggggaga gggccgccct cgacgacctt
     1441 gattctcatg gtctgggtgc cctcgtaggg cttgccttcg ccctcggatg tgcacttgaa
     1501 gtggtggttg ttcacggtgc cctccatgta cagcttcatg ggcatgttct ccttaatcag
     1561 ctcgctcacc atcaccacgt tgatgtcgat ctggtccagg gggatgtact cgccctcgct
     1621 ggtgatcctg ctcttggcgg cccttccaaa cattctgata ttgacttcgt cgttaggagg
     1681 ggcattcttc tcgtgattca ggccaaaagg atcgtcggct tcgggtttct cctgccctgg
     1741 cagacatgcg gggtcccact tcccttccag cactccccag gcagtgctcc acaggatcac
     1801 tccatatgta tttttgcaca ggccgtcagc cagcaggtac agaatggcgc tcacgttctc
     1861 gtggatcacg cagactccgg tgggtgaaat cagccacacg attgggaaca tagtccaagc
     1921 aataaagaag atcaggcgca gcatcttgat cctcaccacc accttctgtg ccagaggtgt
     1981 cccgacagat tcaaagccat acagccccac ttgaaagatt gtgaagacca cactgtactc
     2041 gaacaccagg caggtggccc caaagaaaaa gaacagccac ttcatatact ctgcgggaga
     2101 aattgtagcg gtgattccca tgaccactct aatgatggaa gcggcctgtg caatgggatt
     2161 cagtgggata cttttgtact tcaccagagc catgttagaa atctgtccca gcatgattgg
     2221 acaggtgcac agccaggagg ccattcggct ccaaggaaag aatctgccgt ccctcatctg
     2281 cagggttccg ttgccagtac tggccagtgc ataagtgata gattcgacaa agggcaggta
     2341 cacggactcc caagggacct tgggagccct gctaatcatg aagattgtct ggcagcctga
     2401 gaccacggcc cacaccagct gcagcagtga gacgaaggtg gagtcgattc tggacatatt
     2461 gtagcactcg gtgtgagttg atgcccattc cccatagacg acctgtgatg ccatggtggc
     2521 ggatcctacc gttcgtataa tgtatgctat acgaagttat aagcttatcg ataatcaacc
     2581 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2641 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2701 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2761 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2821 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2881 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2941 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3001 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3061 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3121 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     3181 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     3241 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     3301 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     3361 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     3421 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     3481 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     3541 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     3601 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     3661 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     3721 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3781 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3841 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3901 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3961 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4021 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4081 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4141 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4201 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4261 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4321 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4381 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4441 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4501 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4561 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4621 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4681 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4741 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4801 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4861 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4921 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4981 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5041 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5101 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5161 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5221 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5281 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5341 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5401 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5461 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5521 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5581 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5641 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5701 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5761 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5821 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5881 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5941 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6001 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6061 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6121 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6181 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6241 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6301 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6361 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6421 cttttgctgg ccttttgctc acatgt
//
