LOCUS Exported 4763 bp ds-DNA circular SYN 05-1-2013 DEFINITION Vector for expression and secretion of N-terminally 3xFLAG-tagged proteins in mammalian cells. ACCESSION . VERSION . KEYWORDS p3xFLAG-CMV-8 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4763) AUTHORS Sigma-Aldrich TITLE Direct Submission FEATURES Location/Qualifiers source 1..4763 /organism="synthetic DNA construct" /lab_host="Mammalian Cells" /mol_type="other DNA" primer_bind 141..157 /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" enhancer 318..697 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 698..901 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" CDS 932..976 /codon_start=1 /product="leader sequence from mouse preprotrypsin" /note="preprotrypsin leader" /translation="MSALLILALVGAAVA" CDS 977..1042 /codon_start=1 /product="three tandem FLAG epitope tags, followed by an enterokinase cleavage site" /note="3xFLAG" /translation="DYKDHDGDYKDHDIDYKDDDDK" misc_feature 1040..1106 /note="MCS" /note="multiple cloning site" polyA_signal 1104..1726 /note="hGH poly(A) signal" /note="human growth hormone polyadenylation signal" rep_origin 1935..2070 /note="SV40 ori" /note="SV40 origin of replication" primer_bind complement(2155..2171) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" rep_origin complement(2557..3145) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3316..4176) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(4177..4281) /gene="bla" /note="AmpR promoter" rep_origin 4308..4763 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaattcac catgtctgca cttctgatcc tagctcttgt 961 tggagctgca gttgctgact acaaagacca tgacggtgat tataaagatc atgacatcga 1021 ttacaaggat gacgatgaca agcttgcggc cgcgaattca tcgatagatc tgatatcggt 1081 accagtcgac tctagaggat cccgggtggc atccctgtga cccctcccca gtgcctctcc 1141 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca 1201 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga 1261 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct 1321 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc 1381 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat 1441 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct 1501 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca 1561 ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga 1621 cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca 1681 ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag 1741 cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 1801 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 1861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 1921 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 1981 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc 2041 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ctcgaggaac 2101 tgaaaaacca gaaagttaat tccctatagt gagtcgtatt aaattcgtaa tcatgtcata 2161 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 2221 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 2281 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 2341 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 2401 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 2461 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 2521 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 2581 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 2641 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 2701 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 2761 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 2821 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 2881 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 2941 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 3001 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 3061 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 3121 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 3181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 3241 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 3301 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 3361 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 3421 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 3481 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 3541 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 3601 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 3661 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 3721 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 3781 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 3841 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 3901 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 3961 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 4021 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 4081 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 4141 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 4201 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 4261 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag 4321 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 4381 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 4441 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 4501 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 4561 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 4621 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc 4681 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa 4741 caaaatatta acgcttacaa ttt //