LOCUS Exported 7302 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV packaging vector for hSyn driven KOR DREADD expression.. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-dF-HA-KORD-IRES-mCitrine SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7302) AUTHORS Vardy E, Robinson JE, Li C, Olsen RH, DiBerto JF, Giguere PM, Sassano FM, Huang XP, Zhu H, Urban DJ, White KL, Rittiner JE, Crowley NA, Pleil KE, Mazzone CM, Mosier PD, Song J, Kash TL, Malanga CJ, Krashes MJ, Roth BL TITLE A New DREADD Facilitates the Multiplexed Chemogenetic Interrogation of Behavior. JOURNAL Neuron. 2015 Apr 29. pii: S0896-6273(15)00289-5. doi: 10.1016/j.neuron.2015.03.065. PUBMED 25937170 REFERENCE 2 (bases 1 to 7302) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7302 /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 175..622 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind 660..693 /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 744..777 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(792..1508) /codon_start=1 /product="enhanced variant of YFP (Heikal et al., 2001)" /label=Citrine /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(834..855) /label=EGFP-C /note="EGFP, forward primer" primer_bind 1183..1202 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1442..1463 /label=EGFP-N /note="EGFP, reverse primer" regulatory 1505..1514 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 1515..2088 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1675..1694) /label=IRES-F /note="IRES internal ribosome entry site, forward primer" primer_bind 1904..1921 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS 2602..2613 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" CDS complement(3247..3273) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" protein_bind complement(3287..3320) /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(3371..3404) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3429..4017 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3482..3502) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3900..3911) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4049..4525 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4248..4267) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4565..4705 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4565..4694 /label=AAV2 ITR rep_origin 4780..5235 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4867..4886) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5077..5098 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5252..5271) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5371..5393 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5431..5449) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5517..5621 /gene="bla" /label=AmpR promoter CDS 5622..6482 /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(5840..5859) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6653..7241 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7142..7161 /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca 181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga 241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg 301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt 361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac 421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc 481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca 541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt 601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa 661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc 721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc 781 gcgccctatc acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac 841 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagttt ggactggtag 901 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg 961 tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc 1021 ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac 1081 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg 1141 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg 1201 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag 1261 tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcaca tcaggccgta gccgaaggtg 1321 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc 1381 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt 1441 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg 1501 ctcaccatgg tggcttatca tcgtgttttt caaaggaaaa ccacgtcccc gtggttcggg 1561 gggcctagac gtttttttaa cctcgactaa acacatgtaa agcatgtgca ccgaggcccc 1621 agatcagatc ccatacaatg gggtaccttc tgggcatcct tcagcccctt gttgaatacg 1681 cttgaggaga gccatttgac tctttccaca actatccaac tcacaacgtg gcactggggt 1741 tgtgccgcct ttgcaggtgt atcttataca cgtggctttt ggccgcagag gcacctgtcg 1801 ccaggtgggg ggttccgctg cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa 1861 gcttccagag gaactgcttc cttcacgaca ttcaacagac cttgcattcc tttggcgaga 1921 ggggaaagac ccctaggaat gctcgtcaag aagacagggc caggtttccg ggccctcaca 1981 ttgccaaaag acggcaatat ggtggaaaat aacatataga caaacgcaca ccggccttat 2041 tccaagcggc ttcggccagt aacgttaggg gggggggagg gagaggggcg atatcgcggc 2101 cgctcatact ggtttattca tcccatcgat gtccctcagg taagcaggat cctgaactgt 2161 atttcggact ctgctagtgc tctgccgctc catcctcatc ttcagtggaa agcagaagtc 2221 ccggaaacac cgcttgaagt tttcatcaag aaaggcgtag agaatgggat tcaggctact 2281 gttggtatag cctaaggcga tgcagaagta atagctggag agagcagctg tgctgtggga 2341 ggtgctcccc agagcctcca ccaggatgaa tatgtgaatg ggagtccagc agacgacgaa 2401 gactgccacc accaccagga ccagtctggt gatcctacgc aggttgcgat ctttctctcg 2461 ggagccagaa aggagccgga cgctcttgag acgcaggatc atcagggtgt agcagacgat 2521 gatgatgagg acagggatca cgaaggcaaa gatgaagacg cagatcttca tgaagaggtc 2581 ccaccaggag tagtcatcat ctgggaactg caaggagcac tcaatgacat cgacgtcttc 2641 cctgactttg gtgcctccaa ggactattgc agagatgcca acagatgacg acagcagcca 2701 gatgcagata ttgatgatct ttgccttcaa gggtgtgcgg aagtccaaag ccttcacggg 2761 gtggcacacg gcaatgtagc ggtccacgct catcatggtc aaggtgaaga tgctggtgaa 2821 catgttgtag taattaatgg aaattactat cttgcacagc acatccccaa aaggccagga 2881 attcatcaag tagaccgtac tctgaaaggg catggttgta gtaactaaag catctgccaa 2941 agccaggtta aatatgtaaa tgttggttgc tgtcttcatc tttgtgtatc ggatgatcac 3001 gaacatgacc agcgagttgc ccaccaagcc cacgacgaac actacggagt agaccgccgt 3061 gatgatgacc gggatggccg gggagatgtg cgcgggctcc agctgcgcgt cctccgagcc 3121 ggcgctgccg ttgctgtcgg gctcggccca gccgggaaac caggcgctgc tgttgggggg 3181 caggcaggcg ctcggggcgc aggtagggcc cggctccccg cggaagatct gaatcgggga 3241 ttccatagcg taatctggaa catcgtatgg gtacatggtg gctagcataa cttcgtataa 3301 agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat 3361 ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc gatatcaagc 3421 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 3481 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 3541 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 3601 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 3661 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 3721 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 3781 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt 3841 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 3901 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 3961 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc 4021 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc 4081 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag 4141 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt 4201 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc 4261 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc 4321 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag 4381 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa 4441 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg 4501 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg 4561 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact 4621 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc 4681 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg 4741 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat 4801 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag 4861 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 4921 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 4981 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt 5041 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 5101 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg 5161 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 5221 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 5281 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 5341 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 5401 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta 5461 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg 5521 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 5581 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 5641 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 5701 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 5761 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 5821 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 5881 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 5941 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 6001 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 6061 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 6121 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 6181 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 6241 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 6301 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 6361 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 6421 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 6481 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 6541 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 6601 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 6661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 6721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 6781 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 6841 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 6901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 6961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 7021 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 7081 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 7141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 7201 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 7261 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt //