LOCUS V025025 6044 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025025 VERSION V025025 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..1534 /label="RGS8(NM_033345)" /note="RGS8(NM_033345)" /gene="RGS8" CDS 1541..1606 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1644..1868 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1914..2342 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2356..2685 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2752..3546 /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 3720..3841 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3890..3906) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3914..3930 /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(3938..3968) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3983..4004 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4292..4877) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5048..5908) /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(5909..6013) /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 atgtggaaca ccttaacccg 961 aagcctctct gaccatccag ttggcaaaga ccctcaggcc atgaggactg gccaaagaca 1021 gaacaaaggg atgaggactc gactgggatg cctgtctcac aagtcagact cgtgtagtga 1081 tttcacagct attcttccag acaaacccaa ccgcgctctc aagagattat cgacagaaga 1141 agctacgagg tgggcagatt cctttgatgt gcttctctct cataagtatg gggtggctgc 1201 attccgtgcc ttcttgaaga cggagttcag tgaggagaac ctggaattct ggttggcctg 1261 tgaggagttc aagaagacca ggtcaactgc aaaactggtc tctaaggccc ataggatctt 1321 tgaggagttt gtggatgtgc aggctccacg ggaggtaaac attgacttcc agacccgaga 1381 agccacgagg aagaacctgc aggagccatc cctgacttgc tttgaccaag cccaaggaaa 1441 agtacacagc ctcatggaga aagactctta ccccaggttc ctgaggtcca aaatgtactt 1501 agatctgctg tcccaaagcc agaggaggct cagtctcgag gactacaaag accatgacgg 1561 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 1621 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 1681 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 1741 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 1801 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 1861 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 1921 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 1981 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2041 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2101 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2161 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2221 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2281 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2341 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2401 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 2461 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 2521 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 2581 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 2641 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 2701 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 2761 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 2821 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 2881 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 2941 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3001 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3061 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3121 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3181 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3241 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 3301 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3361 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3421 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3481 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3541 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 3601 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 3661 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 3721 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 3781 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 3841 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 3901 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 3961 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4021 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4081 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4141 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4201 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4261 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4321 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4381 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4441 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4501 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4561 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 4621 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 4681 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 4741 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 4801 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 4861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 4921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 4981 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5041 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5101 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5161 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5221 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5281 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5341 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5401 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 5461 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 5521 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 5581 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 5641 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 5701 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 5761 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 5821 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 5881 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 5941 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6001 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //