LOCUS       sequence-474420-        6522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tetanus toxin light chain in neurons to inhibit synapse 
            activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6522)
  TITLE     AAV-constructs used for labeling and synapse inactivation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6522)
  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            "Unpublished"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-hSyn-TeTxLC-P2A-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..6522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   609..738
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     polyA_signal    838..959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(875..894)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     929..948
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    984..1572
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1090..1101
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1499..1519
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1598..1624)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(join(1644..2357,2358..2360,2361..2363,
                     2364..2366))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MAVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGI
                     KANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVL
                     LEFVTAAGITLGMDELYK"
     primer_bind     complement(1689..1710)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2038..2057
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2297..2318
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2373..2429)
                     /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"
     promoter        complement(3897..4344)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4393..4533
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4393..4522
                     /label=AAV2 ITR
     rep_origin      4608..5063
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4695..4714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4905..4926
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5080..5099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5199..5221
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5259..5277)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5345..5449
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5450..5518,5519..6310)
                     /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(5668..5687)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6481..6522,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg
      661 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
      721 ccatcactag gggttcctgc ggccgctacc acatttgtag aggttttact tgctttaaaa
      781 aacctcccac atctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
      841 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
      901 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
      961 catgtctgac ggccgcaatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg
     1021 agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag
     1081 gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt
     1141 ggcaacacag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt
     1201 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     1261 cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc
     1321 aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct
     1381 gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa
     1441 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     1501 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     1561 cagaggttga ttatcgataa cggccgttct gcagtcaggc gtagtcaggc acgtcgtaag
     1621 ggtatccgcc gctgtcccca tccttacttg tacagctcgt ccatgccgag agtgatcccg
     1681 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     1741 agcttggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     1801 ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg
     1861 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtcg
     1921 ctaaagtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     1981 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     2041 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     2101 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg
     2161 ccccaggaca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     2221 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     2281 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     2341 agctcctcgc ccttgctcac ggccattcta gatggcccgg gattctcttc gacatcccct
     2401 gcttgtttca acagggagaa gttagtggct ccgcttccgt taacagcgct gctagcagcg
     2461 gtacggttgt acaggttttc acggatgttg gtcggcggga tgattttttt gcacagaccg
     2521 atcagtttgg aaaccagacc ggaaccgtcg acgttcctaa aggcgttagt gttcaccctc
     2581 atgttctggc ccttatactc ggacttcaga tccttggact ctatgttaaa gccctcagta
     2641 tcgttatata tagtatcatc cagcaggtta ggtatcttca caggatcgtg gttcatggaa
     2701 aaatagctta agcgagtctt tatgttaaac ttcttgccca gctctatctc agtaaagcca
     2761 tacattatgg agttatacag tatctgaaac ttatcctcgt tcactatata ctggccgttg
     2821 gaatccttat caaactgata cttctgctga tagatctgtt tgtaggagtc gatgtcgatg
     2881 ttcgggtcgt tgcaggaggt aacctgggac agtttgttag cgatggcttt gtagtcgttc
     2941 agggtttttt cgtacaggtc gtttttgata tcgatggaga tcaggttagc gtcctgacca
     3001 ccgaaggtga agagctcttc agcggagatc gggtaggtgt gctgcatgta gatttcctgt
     3061 ttggacggga tgatttcgtg ggaggaaacc tgcatgccgt acagaccgtg cagaacgtgg
     3121 atcagttcgt gcatcagcag cagagccggg tcctggaagt atttggattt accgatggtc
     3181 agggaggtga tgttttcgat aacgttgtcg aaggtcggta cgtattccgg gcagaaagcc
     3241 atctgcatga tggaaccgaa accgtcacgg cacgggaagt agtttttgtt gtcaacacgc
     3301 agaacgatac cacgaacttc gtttttgttc agaaccggac ccggaccgaa gatgatcagg
     3361 ttggtcagca tagcggattt ggtggtagca ccggacggat cctgttccag caggttgaag
     3421 gaaacggagt tggagttggt gtcgaatttg tccagcaggg agtaggagtt acccaggtac
     3481 gggatagcgt tgatgatttt gtccagcaga gcttcaccag caacgttgtt tttgatacgg
     3541 ttgaacagtt taaccatggt ctgcaggaaa cggtctttgt cggagtcggt acgcaggtag
     3601 ttcgggtcgt agtattcgga agcaccttcg atcagggagg acggcgggtt gaagtcttcc
     3661 ggtttggtac caaattcgta acgttccgga acgatccaga tacggtcggt gattttgaag
     3721 gctttgtagt agatgtctag acctttgcag tacggcggtt ccatcatgat gatggtgtcg
     3781 ttgttaaccg ggtcggagta acggaagttg ttgatggtga tcggcatacc ggtttctcga
     3841 cgcatggtgg ccatctgagt ccggtagcgc tagcggatct gacggttcac taaaccctgc
     3901 gctctcaggc acgacacgac tcctccgctg cccaccgcag actgaggcag cgctgagtcg
     3961 ccggcgccgc agcgcagatg gtcgcgcccg tgccccccta tctcgcgcct cgcgtggtgc
     4021 ggtccggctg ggccggcggc ggcgcggacg cgaccaaggt ggccgggaag gggagtttgc
     4081 gggggaccgg cgagtgacgt cagcgcgcct tcagtgctga ggcggcggtg gcgcgcgccg
     4141 ccaggcgggg gcgaaggcac tgtccgcggt gctgaagctg gcagtgcgca cgcgcctcgc
     4201 cgcatcctgt ttcccctccc cctctctgat aggggatgcg caatttgggg aatgggggtt
     4261 gggtgcttgt ccagtgggtc ggggtcggtc gtcaggtagg cacccccacc ccgcctcatc
     4321 ctggtcctaa aacccacttg cactcatacg cagggccctc tgcagtctag acacacgcgg
     4381 taccgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4441 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4501 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4561 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4621 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4681 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4741 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4801 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4861 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4921 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     4981 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5041 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5101 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5161 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5221 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5281 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5341 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5401 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5461 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5521 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5581 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5641 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5701 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5761 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5821 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5881 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5941 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6001 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6061 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6121 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6181 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6241 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6301 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6361 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6421 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6481 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa ac
//
