LOCUS       sequence-199982-        7642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expressing TVA-mCherry fusion and optimised Rabies Glycoprotein 
            (oG) after CRE-mediated recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7642)
  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 7642)
  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-FLEX-TVAmCherry-2A-oG
FEATURES             Location/Qualifiers
     source          1..7642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      127..582
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(214..233)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     424..445
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(599..618)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     718..740
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(778..796)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        864..968
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(969..1037,1038..1829)
                     /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(1187..1206)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2000..2588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2489..2508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2650..2779
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        2855..3158
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3159..3362
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3313..3333
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3359..3383
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3511..3889
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     protein_bind    complement(3989..4022)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(4033..4066)
                     /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)."
     CDS             complement(5666..5731)
                     /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(5741..6445)
                     /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(6016..6035)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6285..6303
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6880..6889
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    6913..6946
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6957..6990
                     /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)."
     polyA_signal    7038..7514
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(7237..7256)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   join(7554..7642,1..52)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(7554..7642,1..41)
                     /label=AAV2 ITR
ORIGIN
        1 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
       61 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
      121 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      181 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
      241 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      301 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
      361 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      421 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
      481 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
      541 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
      601 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
      661 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      721 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
      781 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
      841 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
      901 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      961 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1021 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1081 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1141 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1201 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1261 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1321 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1381 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1441 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1501 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1561 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1621 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1681 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1741 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1801 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1861 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1921 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1981 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2161 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2221 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2281 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2341 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2401 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2461 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2521 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2581 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2641 ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg
     2701 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     2761 tccatcacta ggggttcctg cggccgcacg cgtctagtta ttaatagtaa tcaattacgg
     2821 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     2881 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2941 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     3001 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     3061 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     3121 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     3181 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     3241 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     3301 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     3361 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     3421 gaagacaccg ggaccgatcc agcctccgcg gattcgaatc ccggccggga acggtgcatt
     3481 ggaacgcgga ttccccgtgc caagagtgac gtaagtaccg cctatagagt ctataggccc
     3541 acaaaaaatg ctttcttctt ttaatatact tttttgttta tcttatttct aatactttcc
     3601 ctaatctctt tctttcaggg caataatgat acaatgtatc atgcctcttt gcaccattct
     3661 aaagaataac agtgataatt tctgggttaa ggcaatagca atatttctgc atataaatat
     3721 ttctgcatat aaattgtaac tgatgtaaga ggtttcatat tgctaatagc agctacaatc
     3781 cagctaccat tctgctttta ttttatggtt gggataaggc tggattattc tgagtccaag
     3841 ctaggccctt ttgctaatca tgttcatacc tcttatcttc ctcccacagc tcctgggcaa
     3901 cgtgctggtc tgtgtgctgg cccatcactt tggcaaagaa ttgggattcg aacatcgatt
     3961 gaattcggta cccttcaggt cgacgtcgat aacttcgtat agcatacatt atacgaagtt
     4021 atccgaatcg caataacttc gtataggata ctttatacga agttatttgg cgcgtcgagg
     4081 caagctttca ttagagccgt gtctcgcccc cagatttatg actttcccaa ctgctgatga
     4141 ttttaccgct ctggggtgtg acagacactt ctctaccggt ccccctcaga ttgtgctgag
     4201 ttggctcgga gcggttcact cgtcgccagc aggtcatcag gaaaatgatc agcatgagag
     4261 cagtcagtgc tccagcggag agcagcacgt acttgcccca attaggcagt ccgagatcga
     4321 cgccactgat ccgttcgtgc acgtcgggca ggtggacctc cacaaaatct tcggcctcgt
     4381 cgccgttttt gaacactgtg cttgggtctg cgagaggatg catcagggga atgacgcttg
     4441 acacgagcag ttccatgtgc tgctggagca gactagactg catctcgggg atcagcacgt
     4501 taccatctgg ccccagaatg ataccattaa agaagacccc gttcacatgt gggtgacatc
     4561 gacccccgac tcgcaggcat cccttactag gaatgatttc attccaggtt ctcacagatt
     4621 tataatgggc gtctgcctcc atcagtgtct tgttaaagat ggtgtatgct tttccgaagc
     4681 cagggacgag cttcctcagg tggctcagcc tcctgaagct cacgctcttg gtagtcataa
     4741 ttgactccag agcatcgaga cattcctccc ttttcttcac gagttcctcg accaccagat
     4801 gttcgatctc atcgcttcgg aagtcgtgca gattcacgag ctgcccaggg gggcaccact
     4861 tagtctcgtt agatgtctgc atagcgaccc aggttccgtc catcagccgg agtcccagca
     4921 cgccacagag tttcagcttg caggcacctt tgaggctctt atacagccct ctttcatcca
     4981 caaatccaca agtctcgctg ccttttgagg ccctcttgcc cctgctgttg gtgaagatgt
     5041 cgcaggacat ccccagcctt ggattctcag gcatccagat tgtataatca tggttggtgg
     5101 aacagtaagt ggagctgacg gccactccgg agcagtttcc gccgggaaac actggactgt
     5161 ggagagatcg gtcgtagggg tccaggtcgg cgacagatgg ggaaatgatg accaggctct
     5221 cttttgtggt cttcacagtg cgcagccaat gatagtctgg gtaaggattg tgcaggcttt
     5281 cctcatagcg ggggtcgcca gccatcttcc agttgtaagc tgcccgacag gcgtctggtg
     5341 ttggccgaaa gtgttttctc ttgaaagttg tggtgacata tcccacaaaa tttgtgtagg
     5401 tttctgcctc ggtgaccaca ccagtgcatg tgaacccgtt catcttaata gcagagatat
     5461 agcccacttt cagctccatg taggagaatc cgctcaggtt ggtacagcct tcgtcctcga
     5521 ccacgagatt gttagggcat gacagatggt gaatatcgat aggactccag gggcccagct
     5581 tgtcggggat agtgtagata gggaatttac caaagcagag tgggaagacg agcagaggga
     5641 cgaagaggag agcctgtggg accatgggcc ctgggttgga ctccacgtct cccgccaact
     5701 tgagaaggtc aaaattcaaa gtctgtttca ctccggatcc cttgtacagc tcgtccatgc
     5761 cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt
     5821 tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct
     5881 tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca
     5941 gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg
     6001 cctcccagcc catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc
     6061 gcagcttcac cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca
     6121 ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca
     6181 gcttcaagta gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga
     6241 actgagggga caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct
     6301 tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt
     6361 ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca
     6421 tgttatcctc ctcgcccttg ctcaccccac cccccttgga tgcgctttca agactgaaga
     6481 tgtcagacct gctttttgct ttggacttcc cccatgcagc gataccaccc acagctacca
     6541 ggcagcacag gagcactgca gtgatcagca tccacatgcg gccgtgattc ctggctggca
     6601 gagcacgtcc aggagcaggg gcagtgggag cctctgtgcc gttgtccgtg ggcaccgcgg
     6661 ggatcgcgct ggtcccgcag ccccactcgt cccgcccgtc gtcgcagtcg gggtgtccgt
     6721 cgcacagcca gtcctgcggg taacactccc cgtgggcacc gggcggctcc gagcagcgga
     6781 actgaccggg ggggcaacgg gacaaagaac cgttaccgga cccgttaccg gtcacgttac
     6841 cgggcagcag cagcagcagc agcgcgggca gcagccgcgc catggtggcg aattcggatc
     6901 cggccggcct tgataacttc gtataatgta tgctatacga agttatccga atcgcaataa
     6961 cttcgtataa agtatcctat acgaagttat cgaagatcat caagcttgcc tcgagcagcg
     7021 ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     7081 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     7141 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     7201 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     7261 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     7321 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     7381 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     7441 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     7501 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc
     7561 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     7621 accaaaggtc gcccgacgcc cg
//
