LOCUS Exported 6082 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express recepter TVA for envA with AAV. ACCESSION . VERSION . KEYWORDS pAAV-EF1a-flex-TVA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6082) AUTHORS Menegas W, Bergan JF, Ogawa SK, Isogai Y, Umadevi Venkataraju K, Osten P, Uchida N, Watabe-Uchida M TITLE Dopamine neurons projecting to the posterior striatum form an anatomically distinct subclass. JOURNAL Elife. 2015 Aug 31;4. doi: 10.7554/eLife.10032. PUBMED 26322384 REFERENCE 2 (bases 1 to 6082) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6082 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 242..1419 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 473..1410 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1367..1387 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1450..1483 /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 1534..1567 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(1587..1610) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" protein_bind complement(2067..2100) /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(2151..2184) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2209..2797 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2262..2282) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2680..2691) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2829..3305 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3028..3047) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3345..3485 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3345..3474 /label=AAV2 ITR rep_origin 3560..4015 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3647..3666) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3857..3878 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4032..4051) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4151..4173 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4211..4229) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4297..4401 /gene="bla" /label=AmpR promoter CDS 4402..5262 /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(4620..4639) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5433..6021 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5922..5941 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt 181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat 241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc 1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga 1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg 1561 aagttatggc gcgctagtgg atcctacttg tcgtcgtcgt ccttgtagtc cttggatgcg 1621 ctttcaagac tgaagatgtc agacctgctt tttgctttgg acttccccca tgcagcgata 1681 ccacccacag ctaccaggca gcacaggagc actgcagtga tcagcatcca catgcggccg 1741 tgattcctgg ctggcagagc acgtccagga gcaggggcag tgggagcctc tgtgccgttg 1801 tccgtgggca ccgcggggat cgcgctggtc ccgcagcccc actcgtcccg cccgtcgtcg 1861 cagtcggggt gtccgtcgca cagccagtcc tgcgggtaac actccccgtg ggcaccgggc 1921 ggctccgagc agcggaactg accggggggg caacgggaca aagaaccgtt accggacccg 1981 ttaccggtca cgttaccggg cagcagcagc agcagcagcg cgggcagcag ccgcgccatg 2041 tcggtaccac cacatcaagc tctagcataa cttcgtataa agtatcctat acgaagttat 2101 ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga ataacttcgt 2161 ataatgtatg ctatacgaag ttatgaattc gatatcaagc ttatcgataa tcaacctctg 2221 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 2281 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 2341 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 2401 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 2461 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 2521 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 2581 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc 2641 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 2701 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 2761 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag 2821 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg 2881 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact 2941 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg 3001 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac 3061 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc 3121 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt 3181 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct 3241 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg 3301 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat 3361 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt 3421 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct 3481 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat 3541 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 3601 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 3661 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 3721 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt 3781 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 3841 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 3901 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 3961 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg 4021 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca 4081 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct 4141 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg 4201 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt 4261 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 4321 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 4381 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt 4441 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat 4501 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag 4561 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg 4621 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata 4681 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat 4741 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc 4801 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg 4861 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac 4921 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact 4981 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa 5041 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct 5101 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc 5161 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga 5221 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac 5281 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 5341 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 5401 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 5461 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 5521 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc 5581 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 5641 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 5701 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 5761 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 5821 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 5881 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 5941 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 6001 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 6061 tgctggcctt ttgctcacat gt //