LOCUS       sequence-129491-        7460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of mCherry, TVA and RabG in human and mouse neurons 
            for rabies mono-synaptic transfer experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7460)
  TITLE     Maletic-Savatic LAB, unpublished
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7460)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-hSynI-mCherry-F2A-TVA-T2A-RabG-WPRE-hGH p...
FEATURES             Location/Qualifiers
     source          1..7460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             671..1378
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(816..834)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1084..1103
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1388..1453
                     /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             1934..1987
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    3587..4175
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3640..3660)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4058..4069)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4207..4683
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4406..4425)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4723..4863
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4723..4852
                     /label=AAV2 ITR
     rep_origin      4938..5393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5025..5044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5235..5256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5410..5429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5529..5551
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5589..5607)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5675..5779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5780..5848,5849..6640)
                     /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(5998..6017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6811..7399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7300..7319
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgacgccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
      721 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
      781 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
      841 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
      901 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
      961 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     1021 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     1081 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     1141 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     1201 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     1261 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     1321 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaaggg
     1381 tagtggtgtg aaacagactt tgaattttga ccttctcaag ttggcgggag acgtggagtc
     1441 caacccaggg cccatggcgc ggctgctgcc cgcgctgctg ctgctgctgc tgcccggtaa
     1501 cgtgaccggt aacgggtccg gtaacggttc tttgtcccgt tgcccccccg gtcagttccg
     1561 ctgctcggag ccgcccggtg cccacgggga gtgttacccg caggactggc tgtgcgacgg
     1621 acaccccgac tgcgacgacg ggcgggacga gtggggctgc gggaccagcg cgatccccgc
     1681 ggtgcccacg gacaacggca cagaggctcc cactgcccct gctcctggac gtgctctgcc
     1741 agccaggaat cacggccgca tgtggatgct gatcactgca gtgctcctgt gctgcctggt
     1801 agctgtgggt ggtatcgctg catgggggaa gtccaaagca aaaagcaggt ctgacatctt
     1861 cagtcttgaa agcgcatcca aggagctgct ggtgcctgac aagagccagg cagacttgtt
     1921 ctccggtagt ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc
     1981 tggcccaatg gttcctcagg ctctcctgtt tgtacccctt ctggtttttc cattgtgttt
     2041 tgggaaattc cctatttaca cgataccaga caagcttggt ccctggagtc cgattgacat
     2101 acatcacctc agctgcccaa acaatttggt agtggaggac gaaggatgca ccaacctgtc
     2161 agggttctcc tacatggaac ttaaagttgg atacatctta gccataaaag tgaacgggtt
     2221 cacttgcaca ggcgttgtga cggaggctga aacctacact aacttcgttg gttatgtcac
     2281 aaccacgttc aaaagaaagc atttccgccc aacaccagat gcatgtagag ccgcgtacaa
     2341 ctggaagatg gccggtgacc ccagatatga agagtctcta cacaatccgt accctgacta
     2401 ccgctggctt cgaactgtaa aaaccaccaa ggagtctctc gttatcatat ctccaagtgt
     2461 ggcagatttg gacccatatg acagatccct tcactcgagg gtcttcccta gcgggaagtg
     2521 ctcaggagta gcggtgtctt ctacctactg ctccactaac cacgattaca ccatttggat
     2581 gcccgagaat ccgagactag ggatgtcttg tgacattttt accaatagta gagggaagag
     2641 agcatccaaa gggagtgaga cttgcggctt tgtagatgaa agaggcctat ataagtcttt
     2701 aaaaggagca tgcaaactca agttatgtgg agttctagga cttagactta tggatggaac
     2761 atgggtctcg atgcaaacat caaatgaaac caaatggtgc cctcccgata agttggtgaa
     2821 cctgcacgac tttcgctcag acgaaattga gcaccttgtt gtagaggagt tggtcaggaa
     2881 gagagaggag tgtctggatg cactagagtc catcatgaca accaagtcag tgagtttcag
     2941 acgtctcagt catttaagaa aacttgtccc tgggtttgga aaagcatata ccatattcaa
     3001 caagaccttg atggaagccg atgctcacta caagtcagtc agaacttgga atgagatcct
     3061 cccttcaaaa gggtgtttaa gagttggggg gaggtgtcat cctcatgtga acggggtgtt
     3121 tttcaatggt ataatattag gacctgacgg caatgtctta atcccagaga tgcaatcatc
     3181 cctcctccag caacatatgg agttgttgga atcctcggtt atcccccttg tgcaccccct
     3241 ggcagacccg tctaccgttt tcaaggacgg tgacgaggct gaggattttg ttgaagttca
     3301 ccttcccgat gtgcacaatc aggtctcagg agttgacttg ggtctcccga actgggggaa
     3361 gtatgtatta ctgagtgcag gggccctgac tgccttgatg ttgataattt tcctgatgac
     3421 atgttgtaga agagtcaatc gatcagaacc tacgcaacac aatctcagag ggacagggag
     3481 ggaggtgtca gtcactcccc aaagcgggaa gatcatatct tcatgggaat cacacaagag
     3541 tgggggtgag accagactgt aagaattcga tatcaagctt atcgataatc aacctctgga
     3601 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3661 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3721 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3781 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3841 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3901 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3961 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     4021 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4081 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4141 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag
     4201 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     4261 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     4321 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     4381 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     4441 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     4501 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     4561 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     4621 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     4681 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4741 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4801 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4861 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4921 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     5281 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5341 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     5401 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5461 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5521 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5581 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5641 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5701 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5761 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5821 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5881 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5941 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6001 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6061 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6121 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6181 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6241 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6301 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6361 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6421 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6481 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6541 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6601 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6661 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6721 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6781 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6841 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6901 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     6961 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7021 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7081 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7141 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7201 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7261 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7321 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7381 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7441 ctggcctttt gctcacatgt
//
