LOCUS V043988 8137 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043988 VERSION V043988 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 964..3627 /label="HMGCR(NM_000859)" /note="HMGCR(NM_000859)" /gene="HMGCR" CDS 3634..3699 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3737..3961 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4007..4435 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4449..4778 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4845..5639 /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 5813..5934 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5983..5999) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6007..6023 /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(6031..6061) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6076..6097 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6385..6970) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7141..8001) /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(8002..8106) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgttgt caagactttt tcgaatgcat ggcctctttg tggcctccca tccctgggaa 1021 gtcatagtgg ggacagtgac actgaccatc tgcatgatgt ccatgaacat gtttactggt 1081 aacaataaga tctgtggttg gaattatgaa tgtccaaagt ttgaagagga tgttttgagc 1141 agtgacatta taattctgac aataacacga tgcatagcca tcctgtatat ttacttccag 1201 ttccagaatt tacgtcaact tggatcaaaa tatattttgg gtattgctgg ccttttcaca 1261 attttctcaa gttttgtatt cagtacagtt gtcattcact tcttagacaa agaattgaca 1321 ggcttgaatg aagctttgcc ctttttccta cttttgattg acctttccag agcaagcaca 1381 ttagcaaagt ttgccctcag ttccaactca caggatgaag taagggaaaa tattgctcgt 1441 ggaatggcaa ttttaggtcc tacgtttacc ctcgatgctc ttgttgaatg tcttgtgatt 1501 ggagttggta ccatgtcagg ggtacgtcag cttgaaatta tgtgctgctt tggctgcatg 1561 tcagttcttg ccaactactt cgtgttcatg actttcttcc cagcttgtgt gtccttggta 1621 ttagagcttt ctcgggaaag ccgcgagggt cgtccaattt ggcagctcag ccattttgcc 1681 cgagttttag aagaagaaga aaataagccg aatcctgtaa ctcagagggt caagatgatt 1741 atgtctctag gcttggttct tgttcatgct cacagtcgct ggatagctga tccttctcct 1801 caaaacagta cagcagatac ttctaaggtt tcattaggac tggatgaaaa tgtgtccaag 1861 agaattgaac caagtgtttc cctctggcag ttttatctct ctaaaatgat cagcatggat 1921 attgaacaag ttattaccct aagtttagct ctccttctgg ctgtcaagta catcttcttt 1981 gaacaaacag agacagaatc tacactctca ttaaaaaacc ctatcacatc tcctgtagtg 2041 acacaaaaga aagtcccaga caattgttgt agacgtgaac ctatgctggt cagaaataac 2101 cagaaatgtg attcagtaga ggaagagaca gggataaacc gagaaagaaa agttgaggtt 2161 ataaaaccct tagtggctga aacagatacc ccaaacagag ctacatttgt ggttggtaac 2221 tcctccttac tcgatacttc atcagtactg gtgacacagg aacctgaaat tgaacttccc 2281 agggaacctc ggcctaatga agaatgtcta cagatacttg ggaatgcaga gaaaggtgca 2341 aaattcctta gtgatgctga gatcatccag ttagtcaatg ctaagcatat cccagcctac 2401 aagttggaaa ctctgatgga aactcatgag cgtggtgtat ctattcgccg acagttactt 2461 tccaagaagc tttcagaacc ttcttctctc cagtacctac cttacaggga ttataattac 2521 tccttggtga tgggagcttg ttgtgagaat gttattggat atatgcccat ccctgttgga 2581 gtggcaggac ccctttgctt agatgaaaaa gaatttcagg ttccaatggc aacaacagaa 2641 ggttgtcttg tggccagcac caatagaggc tgcagagcaa taggtcttgg tggaggtgcc 2701 agcagccgag tccttgcaga tgggatgact cgtggcccag ttgtgcgtct tccacgtgct 2761 tgtgactctg cagaagtgaa agcctggctc gaaacatctg aagggttcgc agtgataaag 2821 gaggcatttg acagcactag cagatttgca cgtctacaga aacttcatac aagtatagct 2881 ggacgcaacc tttatatccg tttccagtcc aggtcagggg atgccatggg gatgaacatg 2941 atttcaaagg gtacagagaa agcactttca aaacttcacg agtatttccc tgaaatgcag 3001 attctagccg ttagtggtaa ctattgtact gacaagaaac ctgctgctat aaattggata 3061 gagggaagag gaaaatctgt tgtttgtgaa gctgtcattc cagccaaggt tgtcagagaa 3121 gtattaaaga ctaccacaga ggctatgatt gaggtcaaca ttaacaagaa tttagtgggc 3181 tctgccatgg ctgggagcat aggaggctac aacgcccatg cagcaaacat tgtcaccgcc 3241 atctacattg cctgtggaca ggatgcagca cagaatgttg gtagttcaaa ctgtattact 3301 ttaatggaag caagtggtcc cacaaatgaa gatttatata tcagctgcac catgccatct 3361 atagagatag gaacggtggg tggtgggacc aacctactac ctcagcaagc ctgtttgcag 3421 atgctaggtg ttcaaggagc atgcaaagat aatcctgggg aaaatgcccg gcagcttgcc 3481 cgaattgtgt gtgggaccgt aatggctggg gaattgtcac ttatggcagc attggcagca 3541 ggacatcttg tcaaaagtca catgattcac aacaggtcga agatcaattt acaagacctc 3601 caaggagctt gcaccaagaa gacagccctc gaggactaca aagaccatga cggtgattat 3661 aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc cgtttaaacc 3721 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 3781 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 3841 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 3901 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 3961 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 4021 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 4081 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 4141 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 4201 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 4261 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 4321 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 4381 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 4441 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 4501 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 4561 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 4621 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 4681 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 4741 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 4801 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 4861 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 4921 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 4981 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 5041 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 5101 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 5161 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 5221 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 5281 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 5341 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 5401 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 5461 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 5521 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 5581 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 5641 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 5701 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 5761 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 5821 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 5881 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 5941 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 6001 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 6061 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 6121 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 6181 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 6241 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 6301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 6361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 6421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 6481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 6541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 6601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 6661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 6721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 6781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 6841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6901 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7021 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 7141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 7201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 7261 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 7741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 8101 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //