LOCUS       sequence-314161-        7077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent tetanus toxin light chain construct with 
            nuclear-localized dTomato fluorophore for conditional silencing of 
            neurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7077)
  AUTHORS   Pashkovski SL, Iurilli G, Brann D, Chicharro D, Drummey K, Franks K,
            Panzeri S, Datta SR
  TITLE     Structure and flexibility in cortical representations of odour 
            space.
  JOURNAL   Nature. 2020 Jul;583(7815):253-258. doi: 10.1038/s41586-020-2451-1. 
            Epub 2020 Jul 1.
  PUBMED    32612230
REFERENCE   2  (bases 1 to 7077)
  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"; doi: 
            "10.1038/s41586-020-2451-1"; journalName: "Nature"; date: "2020-07";
            volume: "583"; issue: "7815"; pages: "253-258"
COMMENT     Sequence Label: pAAV-hSyn-FLEX-TeLC-P2A-dTomato
FEATURES             Location/Qualifiers
     source          1..7077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      475..484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    487..520
                     /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    583..616
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(640..1338)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     complement(723..746)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(921..940)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1190..1208
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1339..1359)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1372..1428)
                     /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"
     protein_bind    complement(2843..2876)
                     /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(2947..2980)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3022..3610
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3075..3095)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3493..3504)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3642..4118
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3841..3860)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4158..4298
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4158..4287
                     /label=AAV2 ITR
     rep_origin      4373..4828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4460..4479)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4670..4691
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4845..4864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4964..4986
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5024..5042)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5110..5214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5215..5283,5284..6075)
                     /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(5433..5452)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6246..6834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6735..6754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6896..7025
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaaccgg ctagaggatc cgccgccacc
      481 atggccataa cttcgtataa agtatcctat acgaagttat atcaaaatag gaagaccaat
      541 gcttcaccat cgacccgaat tgccaagcat caccatcgac ccataacttc gtataatgta
      601 tgctatacga agttatttcg cctgcagggg cgcgccttac ttgtacagct cgtccatgcc
      661 gtacaggaac aggtggtggc ggccctcgga gcgctcgtac tgttccacga tggtgtagtc
      721 ctcgttgtgg gaggtgatgt ccagcttggt gtccacgtag tagtagccgg gcagttgcac
      781 gggcttcttg gccatgtaga tggtcttgaa ctccaccagg tagtggccgc cgtccttcag
      841 cttcagggcc tggtggatct cgcccttcag cacgccgtcg cgggggtaca ggcgctcggt
      901 ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggggg ggaagttggt
      961 gccgcgcatc ttcaccttgt agatcagcgt gccgtcctgc agggaggagt cctgggtcac
     1021 ggtcaccaga ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa
     1081 ggacagcttc ttgtaatcgg ggatgtcggc ggggtgcttc acgtacgcct tggagccgta
     1141 catgaactgg ggggacagga tgtcccaggc gaagggcagg gggccgccct tggtcacctt
     1201 cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa
     1261 ctcgtggccg ttcatggagc cctccatgcg caccttgaag cgcatgaact ctttgatgac
     1321 ctcctcgccc ttgctcacca ccttcctctt cttcttgggg gccattctag atggcccggg
     1381 attctcttcg acatcccctg cttgtttcaa cagggagaag ttagtggctc cgcttccgtt
     1441 aacagcgctg ctagcagcgg tacggttgta caggttttca cggatgttgg tcggcgggat
     1501 gatttttttg cacagaccga tcagtttgga aaccagaccg gaaccgtcga cgttcctaaa
     1561 ggcgttagtg ttcaccctca tgttctggcc cttatactcg gacttcagat ccttggactc
     1621 tatgttaaag ccctcagtat cgttatatat agtatcatcc agcaggttag gtatcttcac
     1681 aggatcgtgg ttcatggaaa aatagcttaa gcgagtcttt atgttaaact tcttgcccag
     1741 ctctatctca gtaaagccat acattatgga gttatacagt atctgaaact tatcctcgtt
     1801 cactatatac tggccgttgg aatccttatc aaactgatac ttctgctgat agatctgttt
     1861 gtaggagtcg atgtcgatgt tcgggtcgtt gcaggaggta acctgggaca gtttgttagc
     1921 gatagctttg tagtcgttca gggttttttc gtacaggtcg tttttgatat cgatggagat
     1981 caggttagcg tcctgaccac cgaaggtgaa gagctcttca gcggagatcg ggtaggtgtg
     2041 ctgcatgtag atttcctgtt tggacgggat gatttcgtgg gaggaaacct gcatgccgta
     2101 cagaccgtgc agaacgtgga tcagttcgtg catcagcagc agagccgggt cctggaagta
     2161 tttggattta ccgatggtca gggaggtgat gttttcgata acgttgtcga aggtcggtac
     2221 gtattccggg cagaaagcca tctgcatgat ggaaccgaaa ccgtcacggc acgggaagta
     2281 gtttttgttg tcaacacgca gaacgatacc acgaacttcg tttttgttca gaaccggacc
     2341 cggaccgaag atgatcaggt tggtcagcat agcggatttg gtggtagcac cggacggatc
     2401 ctgttccagc aggttgaagg aaacggagtt ggagttggtg tcgaatttgt ccagcaggga
     2461 gtaggagtta cccaggtacg ggatagcgtt gatgattttg tccagcagag cttcaccagc
     2521 aacgttgttt ttgatacggt tgaacagttt aaccatggtc tgcaggaaac ggtctttgtc
     2581 ggagtcggta cgcaggtagt tcgggtcgta gtattcggaa gcaccttcga tcagggagga
     2641 cggcgggttg aagtcttccg gtttggtacc aaattcgtaa cgttccggaa cgatccagat
     2701 acggtcggtg attttgaagg ctttgtagta gatgtctaga cctttgcagt acggcggttc
     2761 catcatgatg atggtgtcgt tgttaaccgg gtcggagtaa cggaagttgt tgatggtgat
     2821 cggcataccg gtttctcgac gcataacttc gtataggata ctttatacga agttatcttt
     2881 gggattcttc ctattttgat ccaagcatca ccatcgaccc tctagtccag atctcaccat
     2941 cgacccataa cttcgtatag catacattat acgaagttat gtccctcgaa gaggttcgaa
     3001 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3061 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3121 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3181 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3241 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3301 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3361 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3421 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3481 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3541 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3601 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     3661 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     3721 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     3781 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     3841 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     3901 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3961 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     4021 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     4081 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     4141 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4201 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4261 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4321 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4381 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4441 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4501 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4561 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4621 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4681 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4741 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4801 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4861 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4921 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4981 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     5041 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5101 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5161 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5221 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5281 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5341 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5401 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5461 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5521 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5581 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5641 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5701 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5761 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5821 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5881 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5941 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6001 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6061 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6121 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6181 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6241 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6301 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6361 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6421 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6481 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6541 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6601 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6661 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6721 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6781 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6841 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
     6901 aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg
     6961 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     7021 ttcctgcggc cgcacgcgtt taattaagtg tctagactgc agagggccct gcgtatg
//
