LOCUS Exported 6045 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION To study effects of acetyl-mimetic (hyper-acetylated) tubulin. ACCESSION . VERSION . KEYWORDS pEGFP Tubulin K40Q SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6045) AUTHORS Joo EE, Yamada KM TITLE MYPT1 regulates contractility and microtubule acetylation to modulate integrin adhesions and matrix assembly. JOURNAL Nat Commun. 2014 Mar 25;5:3510. doi: 10.1038/ncomms4510. PUBMED 24667306 REFERENCE 2 (bases 1 to 6045) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6045 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 13..601 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 502..521 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 776..1079 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1080..1283 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1234..1254 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 1322..1331 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1328..2044 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1328..2044 /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(1373..1394) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1634..1653) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1981..2002 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3548..3669 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3585..3604) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3639..3658 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3676..4131) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3813..3834) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4025..4044 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4158..4262 /gene="bla" /label=AmpR promoter promoter 4264..4621 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4287..4307) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4472..4607 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4534..4553 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4656..5450 /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(4710..4729) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5320..5339 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5638..5657) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5682..5729 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 61 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt 721 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta 781 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt 841 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg 901 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta 961 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga 1021 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg 1081 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc 1141 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact 1201 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt 1261 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt 1321 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga 1381 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc 1441 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg 1501 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca 1561 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac 1621 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga 1681 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct 1741 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca 1801 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca 1861 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga 1921 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca 1981 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta 2041 caagtccgga ctcagatctc gagtgcgtga gtgcatctcc atccacgttg gccaggctgg 2101 tgtccagatt ggcaatgcct gctgggagct ctactgcctg gaacacggca tccagcccga 2161 tggccagatg ccaagtgacc agaccattgg gggaggagat gactccttca acaccttctt 2221 cagtgagacg ggcgctggca agcacgtgcc ccgggctgtg tttgtagact tggaacccac 2281 agtcattgat gaagttcgca ctggcaccta ccgccagctc ttccaccctg agcagctcat 2341 cacaggcaag gaagatgctg ccaataacta tgcccgaggg cactacacca ttggcaagga 2401 gatcattgac cttgtgttgg accgaattcg caagctggct gaccagtgca ccggtcttca 2461 gggcttcttg gttttccaca gctttggtgg gggaactggt tctgggttca cctccctgct 2521 catggaacgt ctctcagttg attatggcaa gaagtccaag ctggagttct ccatttaccc 2581 agcaccccag gtttccacag ctgtagttga gccctacaac tccatcctca ccacccacac 2641 caccctggag cactctgatt gtgccttcat ggtagacaat gaggccatct atgacatctg 2701 tcgtagaaac ctcgatatcg agcgcccaac ctacactaac cttaaccgcc ttattagcca 2761 gattgtgtcc tccatcactg cttccctgag atttgatgga gccctgaatg ttgacctgac 2821 agaattccag accaacctgg tgccctaccc ccgcatccac ttccctctgg ccacatatgc 2881 ccctgtcatc tctgctgaga aagcctacca tgaacagctt tctgtagcag agatcaccaa 2941 tgcttgcttt gagccagcca accagatggt gaaatgtgac cctcgccatg gtaaatacat 3001 ggcttgctgc ctgttgtacc gtggtgacgt ggttcccaaa gatgtcaatg ctgccattgc 3061 caccatcaaa accaagcgca gcatccagtt tgtggattgg tgccccactg gcttcaaggt 3121 tggcatcaac taccagcctc ccactgtggt gcctggtgga gacctggcca aggtacagag 3181 agctgtgtgc atgctgagca acaccacagc cattgctgag gcctgggctc gcctggacca 3241 caagtttgac ctgatgtatg ccaagcgtgc ctttgttcac tggtacgtgg gtgaggggat 3301 ggaggaaggc gagttttcag aggcccgtga agatatggct gcccttgaga aggattatga 3361 ggaggttggt gtggattctg ttgaaggaga gggtgaggaa gaaggagagg aatactaagg 3421 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact 3481 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt 3541 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 3601 tttcacaaat aaagcatttt tttcactgta ttctagttgt ggtttgtcca aactcatcaa 3661 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt 3721 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa 3781 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa 3841 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac 3901 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga 3961 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa 4021 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc 4081 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca 4141 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 4201 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 4261 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 4321 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 4381 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 4441 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 4501 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 4561 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 4621 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg 4681 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 4741 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 4801 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt 4861 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 4921 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 4981 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 5041 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 5101 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 5161 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg 5221 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 5281 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 5341 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 5401 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct 5461 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac 5521 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat 5581 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga 5641 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa 5701 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca 5761 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc 5821 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc 5881 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt 5941 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 6001 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagat //