LOCUS       sequence-266854-        7062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent Tetanus toxin light chain construct for conditional 
            silencing of neurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7062)
  AUTHORS   Wang et al
  TITLE     Fan Wang Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7062)
  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            "Unpublished"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-FLEX-TeLC-P2A-EYFP-WPRE
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        183..630
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      657..666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    669..702
                     /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)."
     protein_bind    765..798
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(819..1532,1533..1535,1536..1538))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(864..885)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1213..1232
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1472..1493
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1539..1595)
                     /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"
     protein_bind    complement(3010..3043)
                     /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)."
     protein_bind    complement(3114..3147)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3189..3777
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3242..3262)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3660..3671)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3809..4285
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4008..4027)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4325..4465
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4325..4454
                     /label=AAV2 ITR
     rep_origin      4540..4995
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4627..4646)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4837..4858
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5012..5031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5131..5153
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5191..5209)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5277..5381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5382..5450,5451..6242)
                     /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(5600..5619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6413..7001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6902..6921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaatt aagtgtctag actgcagagg gccctgcgta
      181 tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc
      241 gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca
      301 gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg
      361 cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc
      421 gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc
      481 cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg
      541 cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc
      601 ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggctagagga tccgccgcca
      661 ccatggccat aacttcgtat aaagtatcct atacgaagtt atatcaaaat aggaagacca
      721 atgcttcacc atcgacccga attgccaagc atcaccatcg acccataact tcgtataatg
      781 tatgctatac gaagttattt cgcctgcagg ggcgcgcctt acttgtacag ctcgtccatg
      841 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
      901 ttggggtctt tgctcagggc ggactggtag ctcaggtagt ggttgtcggg cagcagcacg
      961 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc
     1021 tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg
     1081 atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc
     1141 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     1201 acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg
     1261 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     1321 gcgaagcact gcaggccgta gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc
     1381 ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc
     1441 tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     1501 accaccccgg tgaacagctc ctcgcccttg ctcaccattg gcccgggatt ctcttcgaca
     1561 tcccctgctt gtttcaacag ggagaagtta gtggctccgc ttccgttaac agcgctgcta
     1621 gcagcggtac ggttgtacag gttttcacgg atgttggtcg gcgggatgat ttttttgcac
     1681 agaccgatca gtttggaaac cagaccggaa ccgtcgacgt tcctaaaggc gttagtgttc
     1741 accctcatgt tctggccctt atactcggac ttcagatcct tggactctat gttaaagccc
     1801 tcagtatcgt tatatatagt atcatccagc aggttaggta tcttcacagg atcgtggttc
     1861 atggaaaaat agcttaagcg agtctttatg ttaaacttct tgcccagctc tatctcagta
     1921 aagccataca ttatggagtt atacagtatc tgaaacttat cctcgttcac tatatactgg
     1981 ccgttggaat ccttatcaaa ctgatacttc tgctgataga tctgtttgta ggagtcgatg
     2041 tcgatgttcg ggtcgttgca ggaggtaacc tgggacagtt tgttagcgat agctttgtag
     2101 tcgttcaggg ttttttcgta caggtcgttt ttgatatcga tggagatcag gttagcgtcc
     2161 tgaccaccga aggtgaagag ctcttcagcg gagatcgggt aggtgtgctg catgtagatt
     2221 tcctgtttgg acgggatgat ttcgtgggag gaaacctgca tgccgtacag accgtgcaga
     2281 acgtggatca gttcgtgcat cagcagcaga gccgggtcct ggaagtattt ggatttaccg
     2341 atggtcaggg aggtgatgtt ttcgataacg ttgtcgaagg tcggtacgta ttccgggcag
     2401 aaagccatct gcatgatgga accgaaaccg tcacggcacg ggaagtagtt tttgttgtca
     2461 acacgcagaa cgataccacg aacttcgttt ttgttcagaa ccggacccgg accgaagatg
     2521 atcaggttgg tcagcatagc ggatttggtg gtagcaccgg acggatcctg ttccagcagg
     2581 ttgaaggaaa cggagttgga gttggtgtcg aatttgtcca gcagggagta ggagttaccc
     2641 aggtacggga tagcgttgat gattttgtcc agcagagctt caccagcaac gttgtttttg
     2701 atacggttga acagtttaac catggtctgc aggaaacggt ctttgtcgga gtcggtacgc
     2761 aggtagttcg ggtcgtagta ttcggaagca ccttcgatca gggaggacgg cgggttgaag
     2821 tcttccggtt tggtaccaaa ttcgtaacgt tccggaacga tccagatacg gtcggtgatt
     2881 ttgaaggctt tgtagtagat gtctagacct ttgcagtacg gcggttccat catgatgatg
     2941 gtgtcgttgt taaccgggtc ggagtaacgg aagttgttga tggtgatcgg cataccggtt
     3001 tctcgacgca taacttcgta taggatactt tatacgaagt tatctttggg attcttccta
     3061 ttttgatcca agcatcacca tcgaccctct agtccagatc tcaccatcga cccataactt
     3121 cgtatagcat acattatacg aagttatgtc cctcgaagag gttcgaattc gatatcaagc
     3181 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3241 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3301 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3361 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3421 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3481 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3541 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3601 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3661 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3721 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3781 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3841 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3901 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3961 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     4021 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     4081 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4141 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4201 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4261 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4321 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4381 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4441 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4501 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4561 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4621 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4681 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4741 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4801 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4861 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4921 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4981 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5041 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5101 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5161 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5221 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5281 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5341 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5401 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5461 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5521 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5581 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5641 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5701 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5761 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5821 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5881 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5941 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6001 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6061 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6121 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6181 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6241 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6301 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6361 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6541 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6781 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
