LOCUS V033491 6596 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033491 VERSION V033491 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..2086 /label="MAPKAPK3(NM_004635)" /note="MAPKAPK3(NM_004635)" /gene="MAPKAPK3" CDS 2093..2158 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2196..2420 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2466..2894 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2908..3237 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3304..4098 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4272..4393 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4442..4458) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4466..4482 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(4490..4520) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4535..4556 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4844..5429) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5600..6460) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(6461..6565) /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 gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggatggtg aaacagcaga 961 ggagcagggg ggccctgtgc ccccgccagt tgcacccggc ggacccggct tgggcggtgc 1021 tccggggggg cggcgggagc ccaagaagta cgcagtgacc gacgactacc agttgtccaa 1081 gcaggtgctg ggcctgggtg tgaacggcaa agtgctggag tgcttccatc ggcgcactgg 1141 acagaagtgt gccctgaagc tcctgtatga cagccccaag gcccggcagg aggtagacca 1201 tcactggcag gcttctggcg gcccccatat tgtctgcatc ctggatgtgt atgagaacat 1261 gcaccatggc aagcgctgtc tcctcatcat catggaatgc atggaaggtg gtgagttgtt 1321 cagcaggatt caggagcgtg gcgaccaggc tttcactgag agagaagctg cagagataat 1381 gcgggatatt ggcactgcca tccagtttct gcacagccat aacattgccc accgagatgt 1441 caagcctgaa aacctactct acacatctaa ggagaaagac gcagtgctta agctcaccga 1501 ttttggcttt gctaaggaga ccacccaaaa tgccctgcag acaccctgct atactcccta 1561 ttatgtggcc cctgaggtcc tgggtccaga gaagtatgac aagtcatgtg acatgtggtc 1621 cctgggtgtc atcatgtaca tcctcctttg tggcttccca cccttctact ccaacacggg 1681 ccaggccatc tccccgggga tgaagaggag gattcgcctg ggccagtacg gcttccccaa 1741 tcctgagtgg tcagaagtct ctgaggatgc caagcagctg atccgcctcc tgttgaagac 1801 agaccccaca gagaggctga ccatcactca gttcatgaac cacccctgga tcaaccaatc 1861 gatggtagtg ccacagaccc cactccacac ggcccgagtg ctgcaggagg acaaagacca 1921 ctgggacgaa gtcaaggagg agatgaccag tgccttggcc actatgcggg tagactacga 1981 ccaggtgaag atcaaggacc tgaagacctc taacaaccgg ctcctcaaca agaggagaaa 2041 aaagcaggca ggcagctcct ctgcctcaca gggctgcaac aaccagctcg aggactacaa 2101 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 2161 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 2221 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 2281 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2341 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 2401 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 2461 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2521 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2581 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2641 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2701 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2761 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 2821 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 2881 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3061 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3121 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3181 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3241 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3301 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3361 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3421 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3481 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3541 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3601 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3661 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3721 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3781 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 3841 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 3901 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 3961 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4021 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4081 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4141 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4201 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4261 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4321 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 4381 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 4441 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4501 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4681 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 5041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 5161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 5221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 5281 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 5341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc 5401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 5821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 6241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 6301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //