LOCUS       sequence-200026-        7395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV constitutively expressing TVA-mCherry fusion and optimised 
            Rabies Glycoprotein (oG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7395)
  AUTHORS   Ciabatti E, Gonzalez-Rueda A, Mariotti L, Morgese F, Tripodi M
  TITLE     Life-Long Genetic and Functional Access to Neural Circuits Using 
            Self-Inactivating Rabies Virus.
  JOURNAL   Cell. 2017 Jul 1. pii: S0092-8674(17)30697-9. doi: 
            10.1016/j.cell.2017.06.014.
  PUBMED    28689641
REFERENCE   2  (bases 1 to 7395)
  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: "Cell. 2017 
            Jul 1. pii: S0092-8674(17)30697-9. doi: 10.1016/j.cell.2017.06.014."
COMMENT     Sequence Label: pAAV-CMV-TVAmCherry-2A-oG
FEATURES             Location/Qualifiers
     source          1..7395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     205..224
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   386..526
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   397..526
                     /label=AAV2 ITR
     polyA_signal    566..1042
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     824..843
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2643..2708)
                     /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"
     CDS             complement(2718..3422)
                     /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(2993..3012)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3262..3280
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3857..3866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3944..4322
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     primer_bind     complement(4450..4474)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4471..4674)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4500..4520)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4675..4978
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   5054..5183
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5245..5833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5325..5344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6004..6795,6796..6864))
                     /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     6627..6646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6865..6969)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7037..7055
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7093..7115)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7215..7234
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(7251..7395,1..311))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(join(7388..7395,1..14))
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
       61 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
      121 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
      181 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct
      241 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
      301 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta
      361 aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag
      421 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
      481 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc
      541 cgcacgtggt tacctacaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta
      601 atcccagcaa tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca
      661 gcctggccaa tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg
      721 tcatgcatgc ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc
      781 aggaggcgga gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat
      841 agaccccgca ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc
      901 tccaccccat aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta
      961 ttaggacaag gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg
     1021 aggggtcaca gggatgccac ccgtagatct ctcgaggcaa gctttcatta gagccgtgtc
     1081 tcgcccccag atttatgact ttcccaactg ctgatgattt taccgctctg gggtgtgaca
     1141 gacacttctc taccggtccc cctcagattg tgctgagttg gctcggagcg gttcactcgt
     1201 cgccagcagg tcatcaggaa aatgatcagc atgagagcag tcagtgctcc agcggagagc
     1261 agcacgtact tgccccaatt aggcagtccg agatcgacgc cactgatccg ttcgtgcacg
     1321 tcgggcaggt ggacctccac aaaatcttcg gcctcgtcgc cgtttttgaa cactgtgctt
     1381 gggtctgcga gaggatgcat caggggaatg acgcttgaca cgagcagttc catgtgctgc
     1441 tggagcagac tagactgcat ctcggggatc agcacgttac catctggccc cagaatgata
     1501 ccattaaaga agaccccgtt cacatgtggg tgacatcgac ccccgactcg caggcatccc
     1561 ttactaggaa tgatttcatt ccaggttctc acagatttat aatgggcgtc tgcctccatc
     1621 agtgtcttgt taaagatggt gtatgctttt ccgaagccag ggacgagctt cctcaggtgg
     1681 ctcagcctcc tgaagctcac gctcttggta gtcataattg actccagagc atcgagacat
     1741 tcctcccttt tcttcacgag ttcctcgacc accagatgtt cgatctcatc gcttcggaag
     1801 tcgtgcagat tcacgagctg cccagggggg caccacttag tctcgttaga tgtctgcata
     1861 gcgacccagg ttccgtccat cagccggagt cccagcacgc cacagagttt cagcttgcag
     1921 gcacctttga ggctcttata cagccctctt tcatccacaa atccacaagt ctcgctgcct
     1981 tttgaggccc tcttgcccct gctgttggtg aagatgtcgc aggacatccc cagccttgga
     2041 ttctcaggca tccagattgt ataatcatgg ttggtggaac agtaagtgga gctgacggcc
     2101 actccggagc agtttccgcc gggaaacact ggactgtgga gagatcggtc gtaggggtcc
     2161 aggtcggcga cagatgggga aatgatgacc aggctctctt ttgtggtctt cacagtgcgc
     2221 agccaatgat agtctgggta aggattgtgc aggctttcct catagcgggg gtcgccagcc
     2281 atcttccagt tgtaagctgc ccgacaggcg tctggtgttg gccgaaagtg ttttctcttg
     2341 aaagttgtgg tgacatatcc cacaaaattt gtgtaggttt ctgcctcggt gaccacacca
     2401 gtgcatgtga acccgttcat cttaatagca gagatatagc ccactttcag ctccatgtag
     2461 gagaatccgc tcaggttggt acagccttcg tcctcgacca cgagattgtt agggcatgac
     2521 agatggtgaa tatcgatagg actccagggg cccagcttgt cggggatagt gtagataggg
     2581 aatttaccaa agcagagtgg gaagacgagc agagggacga agaggagagc ctgtgggacc
     2641 atgggccctg ggttggactc cacgtctccc gccaacttga gaaggtcaaa attcaaagtc
     2701 tgtttcactc cggatccctt gtacagctcg tccatgccgc cggtggagtg gcggccctcg
     2761 gcgcgttcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccaacttg
     2821 atgttgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta ggtggtcttg
     2881 acctcagcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat ctcgcccttc
     2941 agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat ggtcttcttc
     3001 tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt gtagatgaac
     3061 tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc
     3121 atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc ggggatgtcg
     3181 gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag gatgtcccag
     3241 gcgaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag
     3301 gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg
     3361 tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctc
     3421 accccacccc ccttggatgc gctttcaaga ctgaagatgt cagacctgct ttttgctttg
     3481 gacttccccc atgcagcgat accacccaca gctaccaggc agcacaggag cactgcagtg
     3541 atcagcatcc acatgcggcc gtgattcctg gctggcagag cacgtccagg agcaggggca
     3601 gtgggagcct ctgtgccgtt gtccgtgggc accgcgggga tcgcgctggt cccgcagccc
     3661 cactcgtccc gcccgtcgtc gcagtcgggg tgtccgtcgc acagccagtc ctgcgggtaa
     3721 cactccccgt gggcaccggg cggctccgag cagcggaact gaccgggggg gcaacgggac
     3781 aaagaaccgt taccggaccc gttaccggtc acgttaccgg gcagcagcag cagcagcagc
     3841 gcgggcagca gccgcgccat ggtggcgaat tcaatcgatg ttcgaatccc aattctttgc
     3901 caaagtgatg ggccagcaca cagaccagca cgttgcccag gagctgtggg aggaagataa
     3961 gaggtatgaa catgattagc aaaagggcct agcttggact cagaataatc cagccttatc
     4021 ccaaccataa aataaaagca gaatggtagc tggattgtag ctgctattag caatatgaaa
     4081 cctcttacat cagttacaat ttatatgcag aaatatttat atgcagaaat attgctattg
     4141 ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaggc atgatacatt
     4201 gtatcattat tgccctgaaa gaaagagatt agggaaagta ttagaaataa gataaacaaa
     4261 aaagtatatt aaaagaagaa agcatttttt gtgggcctat agactctata ggcggtactt
     4321 acgtcactct tggcacgggg aatccgcgtt ccaatgcacc gttcccggcc gggattcgaa
     4381 tccgcggagg ctggatcggt cccggtgtct tctatggagg tcaaaacagc gtggatggcg
     4441 tctccaggcg atctgacggt tcactaaacg agctctgctt atatagacct cccaccgtac
     4501 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     4561 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     4621 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     4681 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     4741 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     4801 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     4861 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     4921 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     4981 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactaga
     5041 cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5101 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga
     5161 gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5221 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5281 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5341 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5401 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5461 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5581 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5641 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5701 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5761 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5821 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5881 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5941 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6001 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6061 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6121 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6181 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6241 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6301 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6361 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6421 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6481 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6541 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6601 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6661 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6721 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6781 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6841 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6901 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6961 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7021 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     7081 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     7141 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     7201 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa
     7261 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     7321 atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag
     7381 tgttgttcca gtttg
//
