LOCUS Exported 6486 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCDNA3 Flag p38 alpha SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6486) AUTHORS Enslen H, Raingeaud J, Davis RJ TITLE Selective activation of p38 mitogen-activated protein (MAP) kinase isoforms by the MAP kinase kinases MKK3 and MKK6. JOURNAL J Biol Chem. 1998 Jan 16. 273(3):1741-8. PUBMED 9430721 REFERENCE 2 (bases 1 to 6486) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6486 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(63..80) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(234..822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(314..333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(993..1853) /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 1616..1635 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1854..1958) /gene="bla" /label=AmpR promoter primer_bind complement(2033..2052) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2224..2603 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2604..2807 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2758..2778 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2852..2871 /label=T7 /note="T7 promoter, forward primer" promoter 2852..2870 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2898..2921 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(4028..4046) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4029..4046) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4066..4083) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4072..4296 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4342..4770 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4429..4448) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4639..4660 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4779..4799) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4784..5113 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4964..5099 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5026..5045 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5180..5974 /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(5234..5253) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5844..5863 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6148..6269 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6185..6204) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6239..6258 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6318..6334) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6318..6334) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6331..6353) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6342..6358 /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(6366..6396) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6411..6432 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 2881 cttcggcagc caacatggac tataaggacg atgatgacaa aatgtcgcag gagaggccca 2941 cgttctaccg gcaggagctg aacaagacca tctgggaggt gcccgaacga taccagaacc 3001 tgtccccggt gggctcgggc gcctatggct cggtgtgtgc tgcttttgat acaaagacgg 3061 ggcatcgtgt ggcagttaag aagctgtcga gaccgtttca gtccatcatt cacgccaaaa 3121 ggacctaccg agagttgcgt ctgctgaagc acatgaaaca cgaaaatgtg attggtctgt 3181 tggatgtgtt cacacccgca aggtcactgg aggaattcaa tgacgtgtac ctggtgaccc 3241 atctcatggg ggcggacctg aacaacatcg tgaagtgcca gaagctgacc gacgaccacg 3301 ttcagtttct catctaccag atcctccgag ggctgaagta tatacattcg gctgacataa 3361 ttcacaggga cctaaagccc agcaacctag ctgtgaacga agactgtgag ctcaagattc 3421 tggattttgg gctggctcgg cacactgatg atgagatgac aggctacgtg gctaccaggt 3481 ggtaccgagc cccagagatc atgctgaatt ggatgcacta taaccagaca gtggatattt 3541 ggtccgtggg ctgcatcatg gctgagctgt tgaccggaag aacgttgttt cctggtacag 3601 accatattga tcagttgaag ctcattttaa gactcgttgg aaccccaggg gctgagcttc 3661 tgaagaaaat ctcctcagag tctgcaagaa actacattca gtctctggcc cagatgccga 3721 agatgaactt cgcaaatgta tttattggtg ccaatcccct ggctgtcgac ctactggaga 3781 agatgctcgt tttggactca gataagagga tcacagcagc ccaagctctt gcgcatgcct 3841 actttgctca gtaccacgac cctgatgatg agcctgttgc tgacccttat gaccagtcct 3901 ttgaaagcag ggaccttctc atagatgagt ggaagagcct gacctatgat gaagtcatca 3961 gctttgtgcc accacccctt gaccaagaag aaatggagtc ctgagcacct ggtctagagg 4021 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt 4081 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg 4141 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt 4201 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca 4261 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct 4321 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 4381 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt 4441 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc 4501 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg 4561 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg 4621 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct 4681 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg 4741 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg 4801 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 4861 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 4921 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 4981 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 5041 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 5101 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac 5161 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 5221 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 5281 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 5341 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 5401 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 5461 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 5521 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 5581 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 5641 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 5701 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 5761 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 5821 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 5881 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 5941 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 6001 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 6061 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 6121 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 6181 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 6241 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 6301 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 6361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 6421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 6481 cagctg //