LOCUS Exported 7062 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre-dependent Tetanus toxin light chain construct for conditional silencing of neurons.. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-FLEX-TeLC-P2A-EYFP-WPRE 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ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7062) AUTHORS Wang et al TITLE Fan Wang Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 7062) AUTHORS .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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7062 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(669..702) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(765..798) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(819..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 3010..3043 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 3114..3147 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." 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 5382..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 //