LOCUS       sequence-349084-        7138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for mapping synaptic projections of infected neurons; 
            expresses tdTomato to reveal neuronal soma and axons, and expresses 
            EGFP fused to Synaptobrevin-2 to track synaptic terminals. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7138)
  AUTHORS   Li E, Guo J, Oh SJ, Luo Y, Oliveros HC, Du W, Arano R, Kim Y, Chen 
            YT, Eitson J, Lin DT, Li Y, Roberts T, Schoggins JW, Xu W
  TITLE     Anterograde transneuronal tracing and genetic control with 
            engineered yellow fever vaccine YFV-17D.
  JOURNAL   Nat Methods. 2021 Nov 25. pii: 10.1038/s41592-021-01319-9. doi: 
            10.1038/s41592-021-01319-9.
  PUBMED    34824475
REFERENCE   2  (bases 1 to 7138)
  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: "Nat 
            Methods. 2021 Nov 25. pii: 10.1038/s41592-021-01319-9. doi: 
            10.1038/s41592-021-01319-9."
COMMENT     Sequence Label: pAAV-SynaptoTAG2
FEATURES             Location/Qualifiers
     source          1..7138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   49..178
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        214..661
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      664..673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      685..694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             700..2127
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             2137..2193
                     /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"
     CDS             join(2200..2202,2203..2205,2206..2916)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2200..2202,2203..2205,2206..2916)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2245..2266)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2506..2525)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2853..2874
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3313..3901
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3366..3386)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3784..3795)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3933..4409
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4132..4151)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4449..4589
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4449..4578
                     /label=AAV2 ITR
     rep_origin      4664..5119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4751..4770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4961..4982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5136..5155)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5255..5277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5315..5333)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5401..5505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5506..5574,5575..6366)
                     /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(5724..5743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6537..7125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7026..7045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
       61 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
      121 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
      181 ggccgcacgc gtctgcagag ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat
      241 gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca agcacccaac
      301 ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca ggatgcggcg
      361 aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg
      421 cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg gtcccccgca
      481 aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca
      541 ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga
      601 ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca
      661 gtcgccacca tgggatccac cggtgccacc atggtcgaga tggtgagcaa gggcgaggag
      721 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
      781 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      841 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
      901 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
      961 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
     1021 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
     1081 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1141 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag
     1201 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc
     1261 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag
     1321 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgggg
     1381 catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga ggacaacaac
     1441 atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     1501 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1561 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     1621 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     1681 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     1741 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     1801 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1861 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     1921 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     1981 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     2041 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     2101 ctgtacggca tggacgagct gtacaaggga agcggagcta ctaacttcag cctgctgaag
     2161 caggctggag acgtggagga gaaccctgga cctcaattca tggtgagcaa gggcgaggag
     2221 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2281 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2341 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2401 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2461 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2521 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2581 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2641 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2701 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2761 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     2821 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2881 gccgggatca ctctcggcat ggacgagctg tacaagggtg gaggtggatc ggctaccgct
     2941 gccaccgtcc cgcctgccgc cccggccggc gagggtggcc cccctgcacc tcctccaaat
     3001 cttaccagta acaggagact gcagcagacc caggcccagg tggatgaggt ggtggacatc
     3061 atgagggtga atgtggacaa ggtcctggag cgagaccaga agctatcgga actggatgat
     3121 cgcgcagatg ccctccaggc aggggcctcc cagtttgaaa caagtgcagc caagctcaag
     3181 cgcaaatact ggtggaaaaa cctcaagatg atgatcatct tgggagtgat ttgcgccatc
     3241 atcctcatca tcatcatcgt ttacttcagc acttaagaat tgagatctga attcgatatc
     3301 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3361 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3421 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3481 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3541 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3601 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3661 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3721 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3781 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3841 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3901 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
     3961 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     4021 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     4081 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     4141 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     4201 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     4261 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     4321 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     4381 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     4441 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4501 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4561 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4621 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4681 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4741 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4801 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4861 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4921 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4981 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5041 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5101 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5161 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5221 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5281 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5341 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5401 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5461 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5521 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5581 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5641 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5701 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5761 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5821 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5881 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5941 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6001 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6061 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6121 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6181 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6241 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6301 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6361 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6421 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6481 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6541 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6601 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6661 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6721 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6781 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6841 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6901 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6961 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7021 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7081 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacg
//
