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 3614..4222 /label=WPRE /ApEinfo_fwdcolor="#666666" /ApEinfo_revcolor="green" misc_feature 1..141 /label=L-ITR /ApEinfo_fwdcolor="#ffd404" /ApEinfo_revcolor="green" misc_feature 4760..4900 /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(3564..3597) /label=FRT /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" misc_feature 3600..3605 /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(3480..3513) /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(3444..3449) /label=BamHI /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="#ff0816" misc_feature 3458..3463 /label=SnaBI /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="cyan" misc_feature 3472..3479 /label=PacI /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" misc_feature 3464..3471 /label=AscI /ApEinfo_fwdcolor="#ff0208" /ApEinfo_revcolor="green" misc_feature 3452..3457 /label=NheI /ApEinfo_fwdcolor="#ffff00" /ApEinfo_revcolor="green" exon complement(1848..2561) /dnas_title="EYFP (no start)" /label=EYFP (no start) /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" gene complement(2571..3443) /note="GtACR2" misc_feature 3428..3443 /note="reverse primer" misc_feature complement(2563..2570) /note="NotI" PCR_primer complement(2571..2589) /dnas_title="CATCAGAATGTTCGGCAGG" /pair="overlap for TS-EYFP" /primer="CATCAGAATGTTCGGCAGG" /current=0 PCR_primer 2571..2589 /dnas_title="CCTGCCGAACATTCTGATG" /primer="CCTGCCGAACATTCTGATG" /set="Set 2" /pair="Pair 1" /current=1 PCR_primer complement(3428..3444) /dnas_title="CATGGCCTCCCAGGTCG" /primer="CATGGCCTCCCAGGTCG" /set="Set 2" /pair="Pair 1" /current=1 PCR_product 2571..3444 /pair="Pair 1" /set="Set 2" /current=1 /score="1.000" PCR_frame 2571..3445 /note="Reading Frame" /set="Set 2" vector join(1..1555,1851..7497) /source="pAAV-EF1a-FRT-FLEX-GtACR2-EYFP-WPRE" /type="Custom cloned vector" insert 1556..1850 /source="Kv2.1+TLCND w linker PCR product" /type="Custom cloned insert" vector complement(join(1625..1652,1655..1656)) /dnas_title="pAAV-EF1a-FRT-FLEX-GtACR2-EYFP-WPRE" /source="pAAV-EF1a-FRT-FLEX-GtACR2-EYFP-WPRE" misc_feature 1623..1652 /note="GS linker" 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 moust (but in paper they used rabbit)" /note="dendritic targeting domain" misc_feature 1561..1568 /note="AscI" misc_feature 1569..1571 /note="stop" PCR_frame complement(1572..1622) /note="Reading Frame" /set="Set 3" exon complement(1653..1847) /dnas_title="Kv2.1C" /vntifkey="61" /label=Kv2.1 motif misc_feature 1555..1560 /note="BsrGI" misc_feature 1850..1855 /note="BsrGI" source 1..7497 /dnas_title="F-FLEX-ACR2-EYFP-Kv2.1-GS-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 ggcgcgcctc aGGAAGATGT CAGCTGGATA GCGAAAACCT CGCCATCTGC AGGTGACTCT 1621 GCagagcctg atcctgaacc gctaccgctt ccaaaccttg tagcttcggg gaagtcggtg 1681 gcgcaggaaa tgaagctatc gatactagac atgcttctca tgtcaatcac gccctcggtc 1741 ctagctggca gaggagccac tggagatggc atggagctca tttccagttc ctcagggggt 1801 ttactctgag gggccatctc cttagtgttc aggatgggct gagactgctt GTACAGCTCG 1861 TCCATGCCGA GAGTGATCCC GGCGGCGGTC ACGAACTCCA GCAGGACCAT GTGATCGCGC 1921 TTCTCGTTGG GGTCTTTGCT CAGGGCGGAC TGGTAGCTCA GGTAGTGGTT GTCGGGCAGC 1981 AGCACGGGGC CGTCGCCGAT GGGGGTGTTC TGCTGGTAGT GGTCGGCGAG CTGCACGCTG 2041 CCGTCCTCGA TGTTGTGGCG GATCTTGAAG TTCACCTTGA TGCCGTTCTT CTGCTTGTCG 2101 GCCATGATAT AGACGTTGTG GCTGTTGTAG TTGTACTCCA GCTTGTGCCC CAGGATGTTG 2161 CCGTCCTCCT TGAAGTCGAT GCCCTTCAGC TCGATGCGGT TCACCAGGGT GTCGCCCTCG 2221 AACTTCACCT CGGCGCGGGT CTTGTAGTTG CCGTCGTCCT TGAAGAAGAT GGTGCGCTCC 2281 TGGACGTAGC CTTCGGGCAT GGCGGACTTG AAGAAGTCGT GCTGCTTCAT GTGGTCGGGG 2341 TAGCGGGCGA AGCACTGCAG GCCGTAGCCG AAGGTGGTCA CGAGGGTGGG CCAGGGCACG 2401 GGCAGCTTGC CGGTGGTGCA GATGAACTTC AGGGTCAGCT TGCCGTAGGT GGCATCGCCC 2461 TCGCCCTCGC CGGACACGCT GAACTTGTGG CCGTTTACGT CGCCGTCCAG CTCGACCAGG 2521 ATGGGCACCA CCCCGGTGAA CAGCTCCTCG CCCTTGCTCA Cggcggccgc CCTGCCGAAC 2581 ATTCTGATGT TGACCTCGTC GTTGGGGGGG GCATTCTTCT CGTGGTTCAG TCCGAAGGGG 2641 TCGTCGGCCT CAGGCTTCTC CTGGCCAGGG AGGCAAGCCG GGTCCCACTT TCCCTCGAGC 2701 ACTCCCCAGG CGGTGGACCA CAGGATCACG CCGTAGGTGT TCTTGCACAG GCCGTCGGCG 2761 AGCAGATACA GGATGGCGCT CACGTTCTCG TGGATCACGC ACACGCCGGT GGGGGAAATC 2821 AGCCACACGA TGGGGAACAT GGTCCAGGCG ATGAAGAAGA TCAGCCTCAG CATCTTGATC 2881 CTCACCACCA CCTTCTGGGC CAGAGGGGTG CCCACGGACT CGAAGCCGTA CAGTCCCACC 2941 TGGAAGATGG TGAACACCAC GCTGTATTCG AAGACCAGGC AGGTAGCGCC GAAGAAGAAG 3001 AACAGCCACT TCATGTACTC GGCGGGGGAG ATGGTAGCGG TAATGCCCAT CACCACCCTG 3061 ATAATGCTGG CGGCCTGGGC GATGGGGTTC AGAGGGATGC TCTTGTACTT CACCAGAGCC 3121 ATGTTGGAGA TCTGGCCCAG CATAATGGGG CATGTGCACA GCCAGCTGGC CATCCTGGAC 3181 CAGGGAAAGA ACCTGCCGTC CCTCATCTGC AGTGTGCCAT TGCCGGTGCT GGCCAGGGCG 3241 TATGTGATGG ACTCCACGAA GGGCAGGTAC ACGCTCTCCC AGGGCACCTT GGGGGCCCTG 3301 CTGATCATGA AGATGGTCTG GCATCCGCTC ACCACGGCCC ACACCAGCTG CAGCAGGCTC 3361 ACGAAGGTGC TGTCGATCCT GCTCATGTTG TAGCACTCTG TGTGGGTGCT GGCCCACTCG 3421 CCGTACACGA CCTGGGAGGC CATGGATCCC GGctagcTAC GTAggcgcgc cTTAATTAAG 3481 AAGTTCCTAT ACTTCTGATA GAATAGGAAC TTCttgcctt aacccagaaa ttatcactgt 3541 tattctttag aatggtgcaa agaGAAGTTC CTATACTTTC TAGAGAATAG GAACTTCCAG 3601 AATTCgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga 3661 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt 3721 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt 3781 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg 3841 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg 3901 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc 3961 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat 4021 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct 4081 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg 4141 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg 4201 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga 4261 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt 4321 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg 4381 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg 4441 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc 4501 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg 4561 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc 4621 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct 4681 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac 4741 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc 4801 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg 4861 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt 4921 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc 4981 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 5041 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 5101 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 5161 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc 5221 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 5281 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga 5341 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 5401 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg 5461 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg 5521 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt 5581 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc 5641 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc 5701 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 5761 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga 5821 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 5881 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag 5941 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 6001 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta 6061 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 6121 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 6181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag 6241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 6301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 6361 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc 6421 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 6481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 6541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 6601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 6661 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa 6721 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 6781 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 6841 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 6901 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 6961 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 7021 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 7081 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 7141 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 7201 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 7261 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 7321 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 7381 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 7441 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt //