LOCUS Exported 6699 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION ITRACKalpha Donor Construct. ACCESSION . VERSION . KEYWORDS CMV-mEGFP-PKCa-C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6699) AUTHORS Colgan LA, Hu M, Misler JA, Parra-Bueno P, Moran CM, Leitges M, Yasuda R TITLE PKCalpha integrates spatiotemporally distinct Ca(2+) and autocrine BDNF signaling to facilitate synaptic plasticity. JOURNAL Nat Neurosci. 2018 Aug;21(8):1027-1037. doi: 10.1038/s41593-018-0184-3. Epub 2018 Jul 16. PUBMED 30013171 REFERENCE 2 (bases 1 to 6699) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6699 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1326 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELY" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3487..3608 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3524..3543) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3578..3597 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3615..4070) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3752..3773) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3964..3983 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4097..4201 /gene="bla" /label=AmpR promoter promoter 4203..4560 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4226..4246) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4411..4546 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4473..4492 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4595..5389 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4649..4668) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5259..5278 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5577..5596) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5621..5668 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5997..6585 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6486..6505 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacagta tggctgacgt cttcccggcc gccgagccgg tggcgccgca ggacgtggcc 1381 aaccgcttcg cccgcaaagg ggcgctgagg cagaagaacg tgcacgaggt gaagaaccac 1441 cgcttcatcg cgcgcttctt caagcagccc accttctgca gccactgcac cgacttcatc 1501 tgggggtttg ggaaacaagg cttccagtgc caagtttgct gttttgtggt tcacaagagg 1561 tgccatgaat ttgttacttt ttcttgtccg ggggcggata aaggacccga cacagatgac 1621 ccgaggagca agcacaagtt caagatccac acgtatggca gccccacctt ctgtgatcac 1681 tgcggctccc tgctgtacgg actcatccac caggggatga aatgtgacac ctgtgatatg 1741 aacgtgcaca agcagtgcgt gatcaacgtg cccagcctct gcgggatgga ccacacggag 1801 aagaggggcc gcatctacct gaaggccgag gtcacggatg aaaagttgca cgtcacagta 1861 cgagacgcga aaaacctaat ccctatggat ccaaatgggc tttcagatcc ttacgtgaag 1921 ctgaagctta ttcctgaccc caagaacgag agcaaacaga aaaccaagac catccgctcg 1981 acgctgaacc cccggtggga cgagtccttc acgttcaaat taaaaccttc tgataaagac 2041 cggcgactgt ccgtggaaat ctgggactgg gatcgaacca cacggaacga cttcatgggg 2101 tccctttcct ttggggtctc ggagctgatg aagatgccgg ccagcggatg gtacaagctg 2161 ctgaaccaag aggagggcga gtactacaac gtgccgatcc ccgaaggcga cgaggaaggc 2221 aatgtggagc tcaggcagaa attcgagaaa gccaagcttg gccctgccgg caacaaagtc 2281 atcagtccct ccgaggacag gagacagcct tccaacaacc tgggcagagt gaagctcacg 2341 gacttcaact tcctcatggt gctgggcaaa ggcagctttg ggaaggtgat gctggccgac 2401 cgaaagggga cagaggagct gtacgccatc aagatcctga agaaggacgt ggtcatccag 2461 gacgacgacg tggagtgcac catggtggag aagcgggtcc tggcgctgct cgacaagccg 2521 ccgttcctga cgcagctgca ctcctgcttc cagacggtgg accggctgta cttcgtcatg 2581 gagtacgtca acggcgggga cctcatgtac cacatccagc aggtcgggaa gttcaaggag 2641 ccgcaagcag tgttctatgc agcagagatt tccatcgggc tgttctttct tcataaaaga 2701 ggaatcattt atcgggacct gaagttagac aacgtcatgc tggactcgga aggacacatt 2761 aagatcgcgg acttcgggat gtgcaaggag cacatgatgg acggcgtcac gaccaggacc 2821 ttctgcggga cccccgacta catcgcccca gagataatcg cctatcagcc gtacgggaag 2881 tccgtggact ggtgggccta cggcgtcctg ttgtacgaga tgttggccgg gcagcctccg 2941 ttcgacggcg aggacgagga cgagctgttc cagtccatca tggagcacaa cgtctcgtac 3001 cccaagtcct tgtccaagga ggccgtgtcc atctgcaaag ggctgatgac caagcacccc 3061 gggaagcggc tgggctgcgg gcccgagggc gagcgcgacg tgcgggagca tgccttcttc 3121 cggaggatcg actgggagaa gctggagaac cgtgagatcc agccaccctt caagcccaaa 3181 gtgtgcggca aaggagcaga gaactttgac aagttcttca cgcgagggca gcctgtcttg 3241 acgccgcccg accagctggt catcgctaac atcgaccagt ctgattttga aggcttctcc 3301 tacgtcaacc cccagttcgt gcaccccatc ctgcagagcg cggtagccgc gggcccggga 3361 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt 3421 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 3481 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 3541 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 3601 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 3661 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 3721 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 3781 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 3841 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 3901 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 3961 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 4021 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac 4081 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 4141 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 4201 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 4261 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 4321 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 4381 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 4441 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 4501 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 4561 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 4621 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 4681 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 4741 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg 4801 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 4861 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 4921 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 4981 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 5041 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 5101 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg 5161 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 5221 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 5281 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 5341 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 5401 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 5461 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 5521 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa 5581 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 5641 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 5701 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 5761 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 5821 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt 5881 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 5941 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 6001 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 6061 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 6121 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 6181 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 6241 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 6301 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 6361 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 6421 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 6481 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 6541 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 6601 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 6661 tcccctgatt ctgtggataa ccgtattacc gccatgcat //