LOCUS Exported 6935 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent reporter for Ca2+/calmodulin-dependent protein kinase II alpha. ACCESSION . VERSION . KEYWORDS pCAG-mEGFP-CaMKIIa SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6935) AUTHORS Chang JY, Nakahata Y, Hayano Y, Yasuda R TITLE Mechanisms of Ca(2+)/calmodulin-dependent kinase II activation in single dendritic spines. JOURNAL Nat Commun. 2019 Jun 25;10(1):2784. doi: 10.1038/s41467-019-10694-z. PUBMED 31239443 REFERENCE 2 (bases 1 to 6935) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6935 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1201..1223 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1289..1308 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1342..1351 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1348..2064 /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 EFVTAAGITLGMDELYK" primer_bind complement(1393..1414) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1654..1673) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2001..2022 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3549..3568) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3614..3669 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3668..3687) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4030..4046) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4030..4046) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4043..4065) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4054..4070 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4078..4108) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4123..4144 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4203..4398 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4249..4384 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4311..4330 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4404..4538 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4441..4460) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4495..4514 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4606..4623) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4777..5365) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4857..4876) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5536..6396) /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 6159..6178 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6397..6501) /gene="bla" /label=AmpR promoter enhancer 6536..6915 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6917..260 /label=chicken beta-actin promoter ORIGIN 1 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt 61 ttaattattt tgtgcagcga tgggggcggg gggggggggg ggggcgcgcg ccgggggggg 121 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga 181 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa 241 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg 301 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg 361 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt 421 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga 481 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc 541 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc 601 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca 661 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg 721 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg 781 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg 841 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg 901 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat 961 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat 1021 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca 1081 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc 1141 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg 1201 ggttcggctt ctggcgtgtg accggcggct ctagctagag cctctgctaa ccatgttcat 1261 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat 1321 tttggcaaag aattcgctag cgccaccatg gtgagcaagg gcgaggagct gttcaccggg 1381 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc 1441 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc 1501 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc 1561 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa 1621 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc 1681 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc 1741 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc 1801 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac 1861 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac 1921 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac 1981 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact 2041 ctcggcatgg acgagctgta caagtccgga ctcagatctc gagctatggc taccatcacc 2101 tgcacccgat tcacggaaga gtaccagctc ttcgaggaac tgggaaaggg agccttctcc 2161 gtggtgcgca ggtgtgtgaa ggtgctggct ggccaggagt atgctgccaa gattatcaac 2221 accaagaagc tctcagccag agatcaccag aagttggaac gcgaggcccg catctgccgc 2281 ttgttgaagc accccaatat cgtccgactc catgacagca tctccgagga ggggcaccac 2341 taccttatct tcgatctggt cactggtggg gagctgttcg aagacattgt ggcccgggag 2401 tattacagtg aggctgatgc cagccactgt atccagcaga tcctggaggc tgtgctacac 2461 tgtcaccaga tgggggtggt gcatcgcgac ctgaagcctg agaatctgtt gctggcttcg 2521 aagctcaagg gtgctgcggt gaagctggca gactttggcc tggccataga ggttgaggga 2581 gagcagcagg catggtttgg gttcgcaggg acacctggat acctctcccc agaagtgctg 2641 cggaaggacc catacgggaa gcctgtggac ctgtgggcct gtggcgtcat cctgtatatc 2701 ttgctggttg ggtatccccc attctgggat gaggaccagc accgcctgta ccagcagatc 2761 aaagctggtg cctacgattt cccatcacca gaatgggaca ccgtcacccc ggaagccaag 2821 gatctgatca ataagatgct gaccatcaac ccgtccaaac gcatcacggc cgctgaggct 2881 ctcaagcacc cctggatctc gcaccgctcc actgtggcct cctgcatgca cagacaggag 2941 accgtggact gcctgaagaa gttcaatgcc aggaggaaac tgaagggagc catcctcacc 3001 actatgctgg ccaccaggaa cttctccgga gggaagagtg gaggaaacaa gaagaatgat 3061 ggcgtgaagg aatcctctga gagcaccaac accaccatcg aggatgaaga caccaaagtg 3121 cgcaaacagg aaattatcaa agtgacagag cagctgatcg aagccataag caatggagac 3181 tttgaatcct acacgaagat gtgcgaccct ggaatgacag cctttgaacc ggaggccctg 3241 gggaacctgg tcgagggcct ggactttcat cgattctatt ttgaaaacct gtggtcccgg 3301 aacagcaagc ccgtgcacac caccatcctg aaccctcaca tccacctgat gggtgacgag 3361 tcagcctgca tcgcctacat ccgcatcact cagtacctgg atgcgggtgg catcccccgc 3421 acggcccagt cagaggagac ccgtgtctgg caccgcaggg atggaaaatg gcagatcgtc 3481 cacttccaca gatctggggc gccctccgtc ctgccccatt gagcggccgc gactctagag 3541 tcgacggatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat 3601 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg 3661 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat 3721 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta 3781 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa 3841 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca 3901 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc 3961 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc 4021 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 4081 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 4141 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat 4201 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 4261 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 4321 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 4381 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa 4441 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 4501 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca 4561 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 4621 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 4681 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 4741 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 4801 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 4861 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 4921 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 4981 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 5041 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 5101 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 5161 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 5221 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 5281 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 5341 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 5401 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 5461 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 5521 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 5581 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 5641 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 5701 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 5761 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 5821 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 5881 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 5941 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 6001 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 6061 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 6121 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 6181 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 6241 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 6301 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 6361 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 6421 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 6481 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgatcgacat 6541 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat 6601 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 6661 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 6721 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 6781 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 6841 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 6901 atcgctatta ccatggtcga ggtgagcccc acgtt //