LOCUS V018877 8066 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018877 VERSION V018877 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..2746 /label="GRK3(NM_005160)" /note="GRK3(NM_005160)" /gene="GRK3" CDS 2753..2818 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2842..3430 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3496..3544 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3746..4174 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4188..4517 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4584..5378 /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 5554..5675 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5724..5740) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5748..5764 /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(5772..5802) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5817..5838 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6126..6714) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6885..7745) /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(7746..7850) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatggcgga cctggaggct gtgctggccg atgtcagtta 721 cctgatggcc atggagaaga gcaaggcgac cccggccgcc cgcgccagca agaggatcgt 781 cctgccggag cccagtatcc ggagtgtgat gcagaagtac cttgcagaga gaaatgaaat 841 aacctttgac aagattttca atcagaaaat tggtttcttg ctatttaaag atttttgttt 901 gaatgaaatt aatgaagctg tacctcaggt gaagttttat gaagagataa aggaatatga 961 aaaacttgat aatgaggaag accgcctttg cagaagtcga caaatttatg atgcctacat 1021 catgaaggaa cttctttcct gttcacatcc tttctcaaag caagctgtag aacacgtaca 1081 aagtcattta tccaagaaac aagtgacatc aactcttttt cagccataca tagaagaaat 1141 ttgtgaaagc cttcgaggtg acatttttca aaaatttatg gaaagtgaca agttcactag 1201 attttgtcag tggaaaaacg ttgaattaaa tatccatttg accatgaatg agttcagtgt 1261 gcataggatt attggacgag gaggattcgg ggaagtttat ggttgcagga aagcagacac 1321 tggaaaaatg tatgcaatga aatgcttaga taagaagagg atcaaaatga aacaaggaga 1381 aacattagcc ttaaatgaaa gaatcatgtt gtctcttgtc agcacaggag actgtccttt 1441 cattgtatgt atgacctatg ccttccatac cccagataaa ctctgcttca tcctggatct 1501 gatgaacggg ggcgatttgc actaccacct ttcacaacac ggtgtgttct ctgagaagga 1561 gatgcggttt tatgccactg aaatcattct gggtctggaa cacatgcaca atcggtttgt 1621 tgtctacaga gatttgaagc cagcaaatat tctcttggat gaacatggac acgcaagaat 1681 atcagatctt ggtcttgcct gcgatttttc caaaaagaag cctcatgcga gtgttggcac 1741 ccatgggtac atggctcccg aggtgctgca gaaggggacg gcctatgaca gcagtgccga 1801 ctggttctcc ctgggctgca tgcttttcaa acttctgaga ggtcacagcc ctttcagaca 1861 acataaaacc aaagacaagc atgaaattga ccgaatgaca ctcaccgtga atgtggaact 1921 tccagacacc ttctctcctg aactgaagtc ccttttggag ggcttgcttc agcgagacgt 1981 tagcaagcgg ctgggctgtc acggaggcgg ctcacaggaa gtaaaagagc acagcttttt 2041 caaaggtgtt gactggcagc atgtctactt acaaaagtac ccaccaccct tgattcctcc 2101 ccggggagaa gtcaatgctg ctgatgcctt tgatattggc tcatttgatg aagaggatac 2161 caaagggatt aagctacttg attgcgacca agaactctac aagaacttcc ctttggtcat 2221 ctctgaacgc tggcagcaag aagtaacgga aacagtttat gaagcagtaa atgcagacac 2281 agataaaatc gaggccagga agagagctaa aaataagcaa cttggccacg aagaagatta 2341 cgctctgggg aaggactgta ttatgcacgg gtacatgctg aaactgggaa acccatttct 2401 gactcagtgg cagcgtcgct atttttacct ctttccaaat agacttgaat ggagaggaga 2461 gggagagtcc cggcaaaatt tactgacaat ggaacagatt ctctctgtgg aagaaactca 2521 aattaaagac aaaaaatgca ttttgttcag aataaaagga gggaaacaat ttgtcttgca 2581 atgtgagagt gatccagagt ttgtgcagtg gaagaaagag ttgaacgaaa ccttcaagga 2641 ggcccagcgg ctattgcgtc gtgccccgaa gttcctcaac aaacctcggt caggtactgt 2701 ggagctccca aagccatccc tctgtcacag aaacagcaac ggcctcctcg aggactacaa 2761 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 2821 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact 2881 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 2941 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 3001 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 3061 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 3121 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 3181 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 3241 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 3301 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 3361 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 3421 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 3481 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt 3541 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 3601 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 3661 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc 3721 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3781 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3841 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3901 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3961 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 4021 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 4081 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa 4141 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4261 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4321 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 4381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 4441 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 4501 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 4561 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 4621 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 4681 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 4741 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 4801 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 4861 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 4921 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 4981 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 5041 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 5101 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 5161 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 5221 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 5281 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 5341 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa 5401 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 5461 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 5521 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 5581 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 5641 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 5701 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 5761 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 5821 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 5881 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 5941 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 6001 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 6061 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 6121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 6181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 6241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 6301 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 6361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 6421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 6481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 6541 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 6601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 6661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 6721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 6781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 6841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 6901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 6961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 7021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 7081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 7141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 7201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 7261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 7321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 7381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 7441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 7501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 7561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 7621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 7681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 7741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 7801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 7861 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc 7921 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt 7981 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg 8041 acaattgcat gaagaatctg cttagg //