LOCUS Exported 6397 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCDNA3-Flag MKK6(glu) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6397) AUTHORS Raingeaud J, Whitmarsh AJ, Barrett T, Derijard B, Davis RJ TITLE MKK3- and MKK6-regulated gene expression is mediated by the p38 mitogen-activated protein kinase signal transduction pathway. JOURNAL Mol Cell Biol. 1996 Mar . 16(3):1247-55. PUBMED 8622669 REFERENCE 2 (bases 1 to 6397) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6397 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 909..932 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(1950..1968) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1951..1968) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1988..2005) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1994..2218 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2264..2692 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2351..2370) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2561..2582 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2701..2721) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2706..3035 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2886..3021 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2948..2967 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3102..3896 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3156..3175) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3766..3785 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4070..4191 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4107..4126) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4161..4180 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4240..4256) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4240..4256) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4253..4275) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4264..4280 /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(4288..4318) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4333..4354 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4471..4488) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4642..5230) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4722..4741) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5401..6261) /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 6024..6043 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6262..6366) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttcggcag 901 ccaacatgga ctataaggac gatgatgaca aatctcagtc gaaaggcaag aagcgaaacc 961 ctggccttaa aattccaaaa gaagcatttg aacaacctca gaccagttcc acaccaccta 1021 gagatttaga ctccaaggct tgcatttcta ttggaaatca gaactttgag gtgaaggcag 1081 atgacctgga gcctataatg gaactgggac gaggtgcgta cggggtggtg gagaagatgc 1141 ggcacgtgcc cagcgggcag atcatggcag tgaagcggat ccgagccaca gtaaatagcc 1201 aggaacagaa acggctactg atggatttgg atatttccat gaggacggtg gactgtccat 1261 tcactgtcac cttttatggc gcactgtttc gggagggtga tgtgtggatc tgcatggagc 1321 tcatggatac atcactagat aaattctaca aacaagttat tgataaaggc cagacaattc 1381 cagaggacat cttagggaaa atagcagttt ctattgtaaa agcattagaa catttacata 1441 gtaagctgtc tgtcattcac agagacgtca agccttctaa tgtactcatc aatgctctcg 1501 gtcaagtgaa gatgtgcgat tttggaatca gtggctactt ggtggacgag gttgctaaag 1561 aaattgatgc aggttgcaaa ccatacatgg cccctgaaag aataaaccca gagctcaacc 1621 agaagggata cagtgtgaag tctgacattt ggagtctggg catcacgatg attgagttgg 1681 ccatccttcg atttccctat gattcatggg gaactccatt tcagcagctc aaacaggtgg 1741 tagaggagcc atcgccacaa ctcccagcag acaagttctc tgcagagttt gttgacttta 1801 cctcacagtg cttaaagaag aattccaaag aacggcctac atacccagag ctaatgcaac 1861 atccattttt caccctacat gaatccaaag gaacagatgt ggcatctttt gtaaaactga 1921 ttcttggaga ctaatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc 1981 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 2041 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 2101 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 2161 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 2221 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 2281 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 2341 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 2401 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 2461 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 2521 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 2581 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 2641 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 2701 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2761 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 2821 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 2881 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 2941 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 3001 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 3061 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3121 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3181 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 3241 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 3301 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 3361 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 3421 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 3481 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 3541 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 3601 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 3661 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 3721 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 3781 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 3841 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 3901 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 3961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 4021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 4081 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 4141 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 4201 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 4261 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4321 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 4381 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 4441 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 4501 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 4561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 4621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 4801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 4861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 4921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 4981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 5041 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5521 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 5701 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 5761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 5821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 5881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 5941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6001 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6361 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //