LOCUS V019291 6706 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019291 VERSION V019291 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" regulatory 913..922 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 928..1017 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" CDS 1051..2259 /label="RPL3(NM_000967)" /note="RPL3(NM_000967)" /gene="RPL3" polyA_signal 2306..2530 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2576..3004 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3018..3347 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3414..4208 /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 4382..4503 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4552..4568) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4576..4592 /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(4600..4630) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4645..4666 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4954..5539) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5710..6570) /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(6571..6675) /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 aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa 961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca 1021 gcggaaaagc ttggtaccga gctcggatcc atgtctcaca gaaagttctc cgctcccaga 1081 catgggtccc tcggcttcct gcctcggaag cgcagcagca ggcatcgtgg gaaggtgaag 1141 agcttcccta aggatgaccc gtccaagccg gtccacctca cagccttcct gggatacaag 1201 gctggcatga ctcacatcgt gcgggaagtc gacaggccgg gatccaaggt gaacaagaag 1261 gaggtggtgg aggctgtgac cattgtagag acaccaccca tggtggttgt gggcattgtg 1321 ggctacgtgg aaacccctcg aggcctccgg accttcaaga ctgtctttgc tgagcacatc 1381 agtgatgaat gcaagaggcg tttctataag aattggcata aatctaagaa gaaggccttt 1441 accaagtact gcaagaaatg gcaggatgag gatggcaaga agcagctgga gaaggacttc 1501 agcagcatga agaagtactg ccaagtcatc cgtgtcattg cccacaccca gatgcgcctg 1561 cttcctctgc gccagaagaa ggcccacctg atggagatcc aggtgaacgg aggcactgtg 1621 gccgagaagc tggactgggc ccgcgagagg cttgagcagc aggtacctgt gaaccaagtg 1681 tttgggcagg atgagatgat cgacgtcatc ggggtgacca agggcaaagg ctacaaaggg 1741 gtcaccagtc gttggcacac caagaagctg ccccgcaaga cccaccgagg cctgcgcaag 1801 gtggcctgta ttggggcatg gcatcctgct cgtgtagcct tctctgtggc acgcgctggg 1861 cagaaaggct accatcaccg cactgagatc aacaagaaga tttataagat tggccagggc 1921 taccttatca aggacggcaa gctgatcaag aacaatgcct ccactgacta tgacctatct 1981 gacaagagca tcaaccctct gggtggcttt gtccactatg gtgaagtgac caatgacttt 2041 gtcatgctga aaggctgtgt ggtgggaacc aagaagcggg tgctcaccct ccgcaagtcc 2101 ttgctggtgc agacgaagcg gcgggctctg gagaagattg accttaagtt cattgacacc 2161 acctccaagt ttggccatgg ccgcttccag accatggagg agaagaaagc attcatggga 2221 ccactgaaga aagaccgaat tgcaaaggaa gaaggagctt aactcgagtc tagagggccc 2281 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 2341 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 2401 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 2461 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 2521 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg 2581 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 2641 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 2701 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 2761 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 2821 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 2881 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 2941 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3001 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 3061 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3361 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3421 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3481 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3541 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 3601 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 3661 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 3721 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3781 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3841 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3901 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 3961 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 4021 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 4081 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 4141 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4201 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 4261 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 4321 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 4381 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 4441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4501 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 4561 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 4621 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 4681 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 4741 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4801 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4861 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4921 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4981 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 5041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 5101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 5161 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 5221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 5281 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5341 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 5401 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 5461 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg 5521 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 5581 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 5641 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 5701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 6301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //