LOCUS Exported 6744 bp ds-DNA circular SYN 02-MAR-2023 DEFINITION Mammalian expression vector for GFP tagged PKCA.. ACCESSION . VERSION . KEYWORDS GFP-PKCA WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6744) AUTHORS Cameron AJ, Escribano C, Saurin AT, Kostelecky B, Parker PJ TITLE PKC maturation is promoted by nucleotide pocket occupation independently of intrinsic kinase activity. JOURNAL Nat Struct Mol Biol. 2009 Jun;16(6):624-30. doi: 10.1038/nsmb.1606. Epub 2009 May 24. PUBMED 19465915 REFERENCE 2 (bases 1 to 6744) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 2, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6744 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 43..837 /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(97..116) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 707..726 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(1025..1044) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 1069..1116 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 1445..2033 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1934..1953 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 2208..2511 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2512..2715 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2666..2686 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 2754..2763 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2760..3476 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2760..3476 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2805..2826) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3066..3085) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3413..3434 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 5679..5800 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5716..5735) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5770..5789 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5807..6262) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5944..5965) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6156..6175 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6289..6393 /gene="bla" /label=AmpR promoter promoter 6395..8 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(6418..6438) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6603..6738 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6665..6684 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 61 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 121 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 181 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg 241 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 301 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 361 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 421 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 481 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 541 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg 601 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 661 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 721 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 781 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 841 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 901 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 961 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc 1021 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa 1081 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg 1141 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt 1201 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg 1261 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac 1321 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 1381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 1441 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 1501 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 1561 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc 1621 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 1681 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 1741 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 1801 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 1861 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 1921 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 1981 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 2041 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 2101 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag 2161 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 2221 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 2281 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 2341 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 2401 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 2461 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg 2521 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc 2581 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa 2641 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc 2701 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca 2761 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 2821 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 2881 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 2941 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 3001 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 3061 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 3121 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 3181 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 3241 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 3301 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 3361 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 3421 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg 3481 gccggactca gatctcgagc tcaagcttcg aattcatggc tgacgtcttc ccggccgccg 3541 agccggcggc gccgcaggac gtggccaacc gcttcgcccg caaaggggcg ctgaggcaga 3601 agaacgtgca cgaggtgaag aaccaccgct tcatcgcgcg cttcttcaag cagcccacct 3661 tctgcagcca ctgcaccgac ttcatctggg ggtttgggaa acaaggcttc cagtgccaag 3721 tttgctgttt tgtggttcac aagaggtgcc atgaatttgt tactttttct tgtccggggg 3781 cggataaagg acccgacaca gatgacccga ggagcaagca caagttcaag atccacacgt 3841 atggcagccc caccttctgt gatcactgcg gctccctgct gtacggactc atccaccagg 3901 ggatgaaatg tgacacctgt gatatgaacg tgcacaagca gtgcgtgatc aacgtgccca 3961 gcctctgcgg gatggaccac acggagaaga ggggccgcat ctacctgaag gccgaggtca 4021 cggatgaaaa gctgcacgtc acagtacgag acgcgaaaaa cctaatccct atggatccaa 4081 atgggctttc agatccttac gtgaagctga agcttattcc tgaccccaag aacgagagca 4141 aacagaaaac caagaccatc cgctcgacgc tgaacccccg gtgggacgag tccttcacgt 4201 tcaaattaaa accttctgat aaagaccggc gactgtccgt ggaaatctgg gactgggatc 4261 gaaccacacg gaacgacttc atggggtccc tttcctttgg ggtctcggag ctgatgaaga 4321 tgccggccag cggatggtac aagctgctga accaagagga gggcgagtac tacaacgtgc 4381 cgatccccga aggcgacgag gaaggcaatg tggagctcag gcagaaattc gagaaagcca 4441 agcttggccc tgccggcaac aaagtcatca gtccctccga ggacaggaga cagccttcca 4501 acaacctggg cagagtgaag ctcacggact tcaacttcct catggtgctg ggcaaaggca 4561 gctttgggaa ggtgatgctg gccgaccgaa aggggacaga ggagctgtac gccatcaaga 4621 tcctgaagaa ggacgtggtc atccaggacg acgacgtgga gtgcaccatg gtggagaagc 4681 gggtcctggc gctgctcgac aagccgccgt tcctgacgca gctgcactcc tgcttccaga 4741 cggtggaccg gctgtacttc gtcatggagt acgtcaacgg cggggacctc atgtaccaca 4801 tccagcaggt cgggaagttc aaggagccgc aagcagtgtt ctatgcagca gagatttcca 4861 tcgggctgtt ctttcttcat aaaagaggaa tcatttatcg ggacctgaag ttagacaacg 4921 tcatgctgga ctcggaagga cacattaaga tcgcggactt cgggatgtgc aaggagcaca 4981 tgatggacgg cgtcacgacc aggaccttct gcgggacccc cgactacatc gccccagaga 5041 taatcgccta tcagccgtac gggaagtccg tggactggtg ggcctacggc gtcctgttgt 5101 acgagatgtt ggccgggcag cctccgttcg acggcgagga cgaggacgag ctgttccagt 5161 ccatcatgga gcacaacgtc tcgtacccca agtccttgtc caaggaggcc gtgtccatct 5221 gcaaagggct gatgaccaag caccccggga agcggctggg ctgcgggccc gagggcgagc 5281 gcgacgtgcg ggagcatgcc ttcttccgga ggatcgactg ggagaagctg gagaaccgtg 5341 agatccagcc acccttcaag cccaaagtgt gcggcaaagg agcagagaac tttgacaagt 5401 tcttcacgcg agggcagcct gtcttgacgc cgcccgacca gctggtcatc gctaacatcg 5461 accagtctga ttttgaaggc ttctcctacg tcaaccccca gttcgtgcac cccatcctgc 5521 agagcgcggt atgaggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa 5581 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc 5641 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata 5701 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 5761 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt 5821 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 5881 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt 5941 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga 6001 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg 6061 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct 6121 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc 6181 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt 6241 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 6301 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 6361 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga 6421 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 6481 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 6541 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 6601 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 6661 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag 6721 gaggcttttt tggaggccta ggct //