LOCUS Exported 6058 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses splitTVA800. ACCESSION . VERSION . KEYWORDS pAAV-CAG-FLEX-splitTVA800 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6058) TITLE Genetic Neuroengineering Group Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 6058) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6058 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" enhancer 134..513 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 515..792 /label=chicken beta-actin promoter intron 793..1809 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1733..1755 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1817..1836 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1871..1880 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(1886..1919) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(1970..2003) /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 2380..2413 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 2464..2497 /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)." misc_feature 2499..3087 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2552..2572) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2970..2981) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3095..3319 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3321..3461 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3321..3450 /label=AAV2 ITR rep_origin 3536..3991 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3623..3642) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3833..3854 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4008..4027) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4127..4149 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4187..4205) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4273..4377 /gene="bla" /label=AmpR promoter CDS 4378..5238 /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(4596..4615) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5409..5997 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5898..5917 /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 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca 181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc 541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta 601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg 661 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg 721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc 781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc 841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac 901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg 961 tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg 1021 gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc 1081 ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg 1141 agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc 1201 gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa 1261 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc 1321 cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg 1381 ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc 1441 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg 1501 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg 1561 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa 1621 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct 1681 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc 1741 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt 1801 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa 1861 ttccggagcc gccaccatgg ttaatataac ttcgtataat gtatgctata cgaagttatg 1921 cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa taacttcgta 1981 taaagtatcc tatacgaagt tatggcgcgc cctatcagtc ccatctcacc agctcacagc 2041 aaaagatccc tgcagtgatc agcatccaca tgcggccgtg attcctggct ggcagagcac 2101 gtccaggagc agggacagtg ggagcctctg tgccgttgtc cgtgggcacc gcgggggtcg 2161 cgctggtccc gcagccccac tcgtcccgcc cgtcgtcgca gtcggggtgt ccgtcgcaca 2221 gccagtcctg cgggtaacac tccccgtggg caccgggcgg ctccgagcag cggaactgac 2281 cgggggggca acgggacaaa gaaccgttac cggacccgtt accggtcacg ttaccgggca 2341 gcagcagcag cagcagcgcg ggcagcagcc gcgcacctaa taacttcgta tagcatacat 2401 tatacgaagt tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa 2461 agaataactt cgtataggat actttatacg aagttatgaa tcaacctctg gattacaaaa 2521 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 2581 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 2641 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 2701 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 2761 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 2821 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 2881 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc 2941 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 3001 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 3061 ggatctccct ttgggccgcc tccccgcctg gatcctgtgc cttctagttg ccagccatct 3121 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 3181 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 3241 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 3301 gatgcggtgg gctctatggc aggaacccct agtgatggag ttggccactc cctctctgcg 3361 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg 3421 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt 3481 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg 3541 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3601 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3661 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 3721 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg 3781 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 3841 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg 3901 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3961 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct 4021 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 4081 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 4141 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 4201 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 4261 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 4321 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 4381 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 4441 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 4501 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 4561 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 4621 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 4681 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 4741 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 4801 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 4861 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 4921 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 4981 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 5041 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 5101 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 5161 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 5221 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 5281 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 5341 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 5401 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 5461 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 5521 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 5581 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 5641 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 5701 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 5761 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 5821 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 5881 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 5941 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 6001 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt //