LOCUS       sequence-391643-        6640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expressing TVA-mCherry fusion and Rabies Glycoprotein (G) of 
            CVS-N2c strain under neuronal-specific promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  AUTHORS   Lee H, Ciabatti E, Gonzalez-Rueda A, Williams E, Nugent F, Mookerjee
            S, Morgese F, Tripodi M
  TITLE     Combining long-term circuit mapping and network transcriptomics with
            SiR-N2c.
  JOURNAL   Nat Methods. 2023 Apr;20(4):580-589. doi: 
            10.1038/s41592-023-01787-1. Epub 2023 Mar 2.
  PUBMED    36864202
REFERENCE   2  (bases 1 to 6640)
  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/s41592-023-01787-1"; journalName: "Nat Methods"; date: 
            "2023-04"; volume: "20"; issue: "4"; pages: "580-589"
COMMENT     Sequence Label: pAAV-hSyn1-TVAmCherry-2A-G(N2c)
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      95..550
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(182..201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     392..413
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(567..586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     686..708
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(746..764)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        832..936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(937..1005,1006..1797)
                     /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(1155..1174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1968..2556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2457..2476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2618..2747
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2783..3230
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3264..3273
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3708..4412
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3850..3868)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4118..4137
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4422..4487
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     primer_bind     complement(6140..6160)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   join(6520..6640,1..20)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(6520..6640,1..9)
                     /label=AAV2 ITR
ORIGIN
        1 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
       61 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
      121 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
      181 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
      241 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
      301 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
      361 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
      421 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
      481 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
      541 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
      601 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
      661 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
      721 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
      781 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
      841 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      901 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      961 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1021 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1081 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1141 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1201 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1261 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1321 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1381 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1441 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     1501 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     1561 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     1621 attgctgata aatctggagc cggtgagcgt gggtcgcgcg gtatcattgc agcactgggg
     1681 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     1741 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1801 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1861 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1921 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1981 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2041 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2101 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2161 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2221 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2281 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2341 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2401 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2461 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2521 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2581 cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc
     2641 tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     2701 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg gccgcacgcg
     2761 tctgcagagg gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg
     2821 ggggtgccta cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc
     2881 aaattgcgca tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg
     2941 cactgccagc ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc
     3001 gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc
     3061 ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc
     3121 gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc
     3181 ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag acgcgtggac
     3241 gggtaccctt caggtcgaaa ttcgccacca tggcgcggct gctgcccgcg ctgctgctgc
     3301 tgctgctgcc cggtaacgtg accggtaacg ggtccggtaa cggttctttg tcccgttgcc
     3361 cccccggtca gttccgctgc tcggagccgc ccggtgccca cggggagtgt tacccgcagg
     3421 actggctgtg cgacggacac cccgactgcg acgacgggcg ggacgagtgg ggctgcggga
     3481 ccagcgcgat ccccgcggtg cccacggaca acggcacaga ggctcccact gcccctgctc
     3541 ctggacgtgc tctgccagcc aggaatcacg gccgcatgtg gatgctgatc actgcagtgc
     3601 tcctgtgctg cctggtagct gtgggtggta tcgctgcatg ggggaagtcc aaagcaaaaa
     3661 gcaggtctga catcttcagt cttgaaagcg catccaaggg gggtggggtg agcaagggcg
     3721 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     3781 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     3841 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     3901 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     3961 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     4021 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     4081 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     4141 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     4201 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     4261 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     4321 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     4381 actccaccgg cggcatggac gagctgtaca agggatccgg agtgaaacag actttgaatt
     4441 ttgaccttct caagttggcg ggagacgtgg agtccaaccc agggcccatg gtgcctcagg
     4501 tgctgctgtt tgtgctgctg ctgggcttca gcctgtgctt cggcaagttc cccatctaca
     4561 ccatccccga cgagctgggc ccttggagcc ctatcgatat ccaccacctg agctgcccca
     4621 acaacctggt ggtggaagat gagggctgca ccaacctgag cgagttcagc tacatggaac
     4681 tgaaagtggg ctacatcagc gccatcaaag tgaacggctt cacctgtacc ggcgtcgtga
     4741 ccgaggccga gacatacacc aacttcgtgg gctacgtgac caccaccttc aagagaaagc
     4801 acttcagacc cacccccgac gcctgcagag ccgcctacaa ttggaagatg gccggcgacc
     4861 ccagatacga ggaaagcctg cacaacccct accccgacta ccactggctg agaaccgtgc
     4921 ggaccaccaa agagagcctg atcatcatca gccccagcgt gaccgacctg gacccctacg
     4981 acaagagcct gcactccaga gtgttccctg gcggcaagtg cagcggcatc accgtgtcta
     5041 gcacctactg cagcaccaac cacgactaca caatctggat gcccgagaac cccagaccca
     5101 gaaccccctg cgacatcttc accaacagca gaggcaagag agccagcaac ggcaacaaga
     5161 cctgcggctt cgtggacgag agaggcctgt acaagtccct gaagggcgcc tgccggctga
     5221 agctgtgtgg cgtgctggga ctgagactga tggacggcac atgggtggcc atgcagacca
     5281 gcgacgagac aaagtggtgc ccccctgatc agctcgtgaa cctgcacgac ttcagatccg
     5341 acgagatcga gcatctggtg gtggaagaac tcgtgaagaa aagggaagag tgcctggacg
     5401 ccctggaatc catcatgacc accaagagcg tgtccttcag aaggctgagc cacctgagaa
     5461 agctggtgcc cggctttggc aaggcttaca ccatcttcaa caagacactg atggaagccg
     5521 acgcccacta caagtctgtg cggacctgga acgagatcat ccccagcaag ggctgcctga
     5581 aagttggagg cagatgccac cctcacgtga acggcgtgtt cttcaacggc atcatcctgg
     5641 gccccgacga ccacgtgctg atccctgaga tgcagagcag cctgctgcag cagcatatgg
     5701 aactgctgga aagcagcgtg atccccctga tgcaccctct ggccgaccct agcaccgtgt
     5761 tcaaagaggg cgacgaggct gaggacttcg tggaagtgca tctgcccgac gtgtacaagc
     5821 agatcagcgg cgtggacctg ggcctgccca actggggcaa atatgtgctg atgaccgctg
     5881 gcgccatgat cggcctggtg ctgatcttct ccctgatgac ctggtgcaga agggccaaca
     5941 gacccgagag caagcagaga agcttcggcg gcacaggcgg caacgtgtcc gtgacaagcc
     6001 agagcggcaa agtgatccct agctgggaga gctacaagag cggcggagag atcagactgt
     6061 gatgatgact cgagaagctt cgagataatc aacctctgga ttacaaaatt tgtgaaagat
     6121 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     6181 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     6241 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6301 ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg
     6361 ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc tttatttgta
     6421 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     6481 gttcaggggg agatgtggga ggttttttaa agcggccgca ggaaccccta gtgatggagt
     6541 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     6601 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
//
