LOCUS pAAV_EF1a_F14FRTFl 7041 bp ds-DNA circular 22-MAY-2015 DEFINITION . ACCESSION VERSION SOURCE . ORGANISM . COMMENT COMMENT ApEinfo:methylated:1 FEATURES Location/Qualifiers misc_feature 3389..3997 /label=WPRE /ApEinfo_fwdcolor="#666666" /ApEinfo_revcolor="green" misc_feature 1..141 /label=L-ITR /ApEinfo_fwdcolor="#ffd404" /ApEinfo_revcolor="green" misc_feature 4535..4675 /label=R-ITR /ApEinfo_fwdcolor="#ffd00a" /ApEinfo_revcolor="green" misc_feature 1544..1549 /label=EcoRI /ApEinfo_fwdcolor="#ff0510" /ApEinfo_revcolor="green" misc_feature 156..1419 /label=EF1a promoter /ApEinfo_fwdcolor="#ff08b6" /ApEinfo_revcolor="green" misc_feature 1420..1425 /label=KpnI /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" Region complement(3339..3372) /label=FRT /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" misc_feature 3375..3380 /label=EcoRI(1) /ApEinfo_label="EcoRI" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" Region 1510..1543 /label=FRT(1) /ApEinfo_label="FRT" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" misc_feature complement(3255..3288) /label=F14 /ApEinfo_fwdcolor="#ffdd07" /ApEinfo_revcolor="#ffdd07" misc_feature 1426..1459 /label=F14(1) /ApEinfo_label="F14" /ApEinfo_fwdcolor="#ffdd07" /ApEinfo_revcolor="#ffdd07" misc_feature complement(3219..3224) /label=BamHI /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="#ff0816" misc_feature 3233..3238 /label=SnaBI /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="cyan" misc_feature 3247..3254 /label=PacI /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" misc_feature 3239..3246 /label=AscI /ApEinfo_fwdcolor="#ff0208" /ApEinfo_revcolor="green" misc_feature 1625..1630 /dnas_title="BsrGI" /label=BsrGI /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" misc_feature 3227..3232 /label=NheI /ApEinfo_fwdcolor="#ffff00" /ApEinfo_revcolor="green" exon complement(1623..2336) /dnas_title="EYFP (no start)" /label=EYFP (no start) /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" gene complement(2346..3218) /note="GtACR2" misc_feature 3203..3218 /note="reverse primer" misc_feature complement(2338..2345) /note="NotI" PCR_primer complement(2346..2364) /pair="overlap for TS-EYFP" /primer="CATCAGAATGTTCGGCAGG" /current=0 PCR_primer 2346..2364 /primer="CCTGCCGAACATTCTGATG" /set="Set 2" /pair="Pair 1" /current=1 PCR_primer complement(3203..3219) /primer="CATGGCCTCCCAGGTCG" /set="Set 2" /pair="Pair 1" /current=1 PCR_product 2346..3219 /pair="Pair 1" /set="Set 2" /current=1 /score="1.000" PCR_frame 2346..3220 /note="Reading Frame" /set="Set 2" vector join(1..1555,1626..7272) /source="pAAV-EF1a-FRT-FLEX-GtACR2-EYFP-WPRE" /type="Custom cloned vector" insert 1556..1625 /source="TLCN PCR insert" /type="Custom cloned insert" misc_feature complement(1572..1622) /dnas_title="TlcnC" /note="TLCND" /note="from paper: A Novel Phenylalanine-Based Targeting Signal Directs Telencephalin to Neuronal Dendrites, Mitsui et al" /translation="AESPADGEVFAIQLTSS" /note="from mouse (but in paper they used rabbit)" /note="dendritic targeting domain from telencephalin" misc_feature 1561..1568 /note="PmeI" misc_feature 1569..1571 /note="stop" misc_feature 1555..1560 /note="BsrGI" misc_feature complement(1665..1686) /note="EGFP-C geneqiz seq primer" source 1..7272 /dnas_title="F-FLEX-ACR2-EYFP-TLCND" 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 GTACCGAAGT TCCTATTCTA 1441 TCAGAAGTAT AGGAACTTCg cagaatggta gctggattgt agctgctatt agcaatatga 1501 aacctcttaG AAGTTCCTAT TCTCTAGAAA GTATAGGAAC TTCgaattcT TACTTGTACA 1561 gtttaaactc aGGAAGATGT CAGCTGGATA GCGAAAACCT CGCCATCTGC AGGTGACTCT 1621 GCctTGTACA GCTCGTCCAT GCCGAGAGTG ATCCCGGCGG CGGTCACGAA CTCCAGCAGG 1681 ACCATGTGAT CGCGCTTCTC GTTGGGGTCT TTGCTCAGGG CGGACTGGTA GCTCAGGTAG 1741 TGGTTGTCGG GCAGCAGCAC GGGGCCGTCG CCGATGGGGG TGTTCTGCTG GTAGTGGTCG 1801 GCGAGCTGCA CGCTGCCGTC CTCGATGTTG TGGCGGATCT TGAAGTTCAC CTTGATGCCG 1861 TTCTTCTGCT TGTCGGCCAT GATATAGACG TTGTGGCTGT TGTAGTTGTA CTCCAGCTTG 1921 TGCCCCAGGA TGTTGCCGTC CTCCTTGAAG TCGATGCCCT TCAGCTCGAT GCGGTTCACC 1981 AGGGTGTCGC CCTCGAACTT CACCTCGGCG CGGGTCTTGT AGTTGCCGTC GTCCTTGAAG 2041 AAGATGGTGC GCTCCTGGAC GTAGCCTTCG GGCATGGCGG ACTTGAAGAA GTCGTGCTGC 2101 TTCATGTGGT CGGGGTAGCG GGCGAAGCAC TGCAGGCCGT AGCCGAAGGT GGTCACGAGG 2161 GTGGGCCAGG GCACGGGCAG CTTGCCGGTG GTGCAGATGA ACTTCAGGGT CAGCTTGCCG 2221 TAGGTGGCAT CGCCCTCGCC CTCGCCGGAC ACGCTGAACT TGTGGCCGTT TACGTCGCCG 2281 TCCAGCTCGA CCAGGATGGG CACCACCCCG GTGAACAGCT CCTCGCCCTT GCTCACggcg 2341 gccgcCCTGC CGAACATTCT GATGTTGACC TCGTCGTTGG GGGGGGCATT CTTCTCGTGG 2401 TTCAGTCCGA AGGGGTCGTC GGCCTCAGGC TTCTCCTGGC CAGGGAGGCA AGCCGGGTCC 2461 CACTTTCCCT CGAGCACTCC CCAGGCGGTG GACCACAGGA TCACGCCGTA GGTGTTCTTG 2521 CACAGGCCGT CGGCGAGCAG ATACAGGATG GCGCTCACGT TCTCGTGGAT CACGCACACG 2581 CCGGTGGGGG AAATCAGCCA CACGATGGGG AACATGGTCC AGGCGATGAA GAAGATCAGC 2641 CTCAGCATCT TGATCCTCAC CACCACCTTC TGGGCCAGAG GGGTGCCCAC GGACTCGAAG 2701 CCGTACAGTC CCACCTGGAA GATGGTGAAC ACCACGCTGT ATTCGAAGAC CAGGCAGGTA 2761 GCGCCGAAGA AGAAGAACAG CCACTTCATG TACTCGGCGG GGGAGATGGT AGCGGTAATG 2821 CCCATCACCA CCCTGATAAT GCTGGCGGCC TGGGCGATGG GGTTCAGAGG GATGCTCTTG 2881 TACTTCACCA GAGCCATGTT GGAGATCTGG CCCAGCATAA TGGGGCATGT GCACAGCCAG 2941 CTGGCCATCC TGGACCAGGG AAAGAACCTG CCGTCCCTCA TCTGCAGTGT GCCATTGCCG 3001 GTGCTGGCCA GGGCGTATGT GATGGACTCC ACGAAGGGCA GGTACACGCT CTCCCAGGGC 3061 ACCTTGGGGG CCCTGCTGAT CATGAAGATG GTCTGGCATC CGCTCACCAC GGCCCACACC 3121 AGCTGCAGCA GGCTCACGAA GGTGCTGTCG ATCCTGCTCA TGTTGTAGCA CTCTGTGTGG 3181 GTGCTGGCCC ACTCGCCGTA CACGACCTGG GAGGCCATGG ATCCCGGcta gcTACGTAgg 3241 cgcgccTTAA TTAAGAAGTT CCTATACTTC TGATAGAATA GGAACTTCtt gccttaaccc 3301 agaaattatc actgttattc tttagaatgg tgcaaagaGA AGTTCCTATA CTTTCTAGAG 3361 AATAGGAACT TCCAGAATTC gatatcaagc ttatcgataa tcaacctctg gattacaaaa 3421 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 3481 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 3541 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 3601 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 3661 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 3721 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 3781 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc 3841 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 3901 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 3961 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg 4021 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt 4081 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt 4141 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac 4201 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct 4261 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt 4321 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg 4381 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg 4441 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat 4501 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc 4561 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc 4621 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc 4681 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc 4741 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 4801 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 4861 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 4921 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac 4981 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 5041 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt 5101 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 5161 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca 5221 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg 5281 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct 5341 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg 5401 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt 5461 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 5521 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 5581 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 5641 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 5701 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 5761 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 5821 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5881 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5941 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 6001 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 6061 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 6121 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 6181 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 6241 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 6301 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 6361 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 6421 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 6481 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 6541 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 6601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 6661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 6721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 6781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 7021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 7081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 7141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 7201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 7261 ttgctcacat gt //