LOCUS V019315 9404 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019315 VERSION V019315 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..4084 /label="Srebf1(NM_001276707)" /note="Srebf1(NM_001276707)" /gene="Srebf1" CDS 4091..4156 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4180..4768 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4834..4882 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5084..5512 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5526..5855 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5922..6716 /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 6892..7013 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7062..7078) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7086..7102 /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(7110..7140) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7155..7176 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7464..8052) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8223..9083) /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(9084..9188) /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 ccatggacga gctacccttc ggtgaggcgg ctctggaaca 721 ggcactggcc gaggtgtgcg aaatggacgc ggcgctcttg accgacatcg aagacatgct 781 tcagctcatc aacaaccaag acagtgactt ccctggccta tttgatgccc cctatgctgg 841 gggtgagaca ggagacacag gccccagcag ccctggtgcc agctctcctg agagcttctc 901 ttctcctgct tctctgggct cctctctgga agccttcctg ggaggaccca aggtgacacc 961 tgcacccttg tcccctccac catcggcacc cactgctgta aagatgtacc cgtccgtgcc 1021 ccccttctcc cctgggcctg gaatcaaaga ggagccagtg ccactcacca tcctgcagcc 1081 cccagcacca cagccatcgc cagggaccct gttgcctccg agcttccctc ctccacctgt 1141 gcagctcagc cctgctcctg tgctggggta ctcaagcctg ccttccggct tctcaggaac 1201 ccttcctggg aacacccagc agacgccatc tagcctgcca ctgggctcca cgccaggaat 1261 ctcgcccacc cccttacaca cccaggtcca gagctcggcc gcccagcagc cgccgccagc 1321 ctcagcagcc cctagaatga gcactgtggc ctcacagatc cagcaggtcc ccgttgtact 1381 gcagccacac ttcatcaagg cagactcgct gctgctgaca gctgtaaaga cagacacagg 1441 agccacaatg aagaccgcag gcatcaacac cctggctcct ggcacagccg tgcaggcagg 1501 ccccttgcag accctggtga gtggagggac catcctggcc acagtaccac tggttgtgga 1561 cacagacaaa ctgcccatcc accgactagc agctggtggc aaggccctgg gctcagctca 1621 gagccgtggt gagaagcgca cagcccacaa tgccattgag aagcgctacc gttcctctat 1681 caatgacaag attgtggagc tcaaggacct ggtggtgggc actgaggcaa agctgaataa 1741 atctgctgtc ttgcgcaagg ccatcgacta catccgcttc ttacagcaca gcaaccagaa 1801 actcaagcag gagaacctga ccctgcgaag tgctcacaaa agcaaatcac tgaaagacct 1861 ggtgtcagct tgtggcagtg gaggaggcac agatgtgtct atggagggca tgaaacctga 1921 agtggtagaa acgctgaccc ctccaccctc agacgccggc tcaccctccc agagtagccc 1981 cttgtccttg ggcagcagag gcagcagcag tggtggcagt gactctgagc ccgacagccc 2041 agcctttgag gataaccagg tgaaagccca gcggctgcct tcacatagcc gaggcatgct 2101 ggaccgctcc cgcctggccc tgtgtgtact ggtcttcctg tgtctgacct gcaacccatt 2161 ggcctcactg tttggctggg gcatcctcac tccctctgat gcttcgggtg tgcaccgtag 2221 ttctgggcgc agcatgctgg aggccgagag cagagatggc tctaattgga cccagtggtt 2281 gctgccaccc ctagtctggc tggccaatgg actactagtg ttggcctgct tggctcttct 2341 ctttgtctac ggggaacctg tgaccaggcc acactccggc ccggctgtac acttctggag 2401 acatcgcaaa caagctgacc tggatttggc ccggggagat tttgcccagg ccgctcaaca 2461 gctgtggctg gccttgcaag ccctgggccg gcccctgccc acctcaaacc tggatctggc 2521 ctgcagcctg ctttggaacc tcgtccgcca cctgctgcag cgtctttggg tgggccgctg 2581 gctggcaggc caggctgggg gcctgcagag ggactacagg ctgagaaagg atgctcgtgc 2641 cagtgcccga gatgcggctg tcgtctacca taagctgcac cagctgcatg ccatgggcaa 2701 gtacacagga ggccatcttg ttgcttctaa cctggcactg agtgccctta acctggctga 2761 gtgtgcagga gatgctatat ccatggcaac actggcagag atctacgtgg cagctgccct 2821 aagggtcaaa accagcctcc ccagagcctt gcacttcttg acacgtttct tcctgagtag 2881 tgcccgccag gcctgcctgg cacagagtgg tgcagtgcct cttgccatgc agtggctctg 2941 ccaccctgta ggtcaccgtt tcttcgtgga tggggactgg gctgtacacg gtgcccccca 3001 ggagagtctg tacagcgtgg ctgggaaccc agtggatcca ctggcccagg tgacccgact 3061 attctgtgaa catctcctgg agcgagcatt gaactgtatc gctcagccca gcccaggggc 3121 agctgatgga gacagggagt tctcagatgc tcttggatat ctacagttgc taaatagctg 3181 ttctgacgct gtcggagctc ctgcgtgcag cttctctgtc agttccagca tggctaccac 3241 cactggcaca gacccagtgg ccaagtggtg ggcctcactg acagccgtgg tgatccactg 3301 gctgaggcgg gatgaggagg cagctgaacg cttataccca ctggtagagc acattcccca 3361 agtgctgcag gaaactgaga gaccccttcc cagggcagct ctgtactcct tcaaggctgc 3421 ccgggctctg ctggaccaca gaaaggtgga atccagccca gccagcctgg ccatctgtga 3481 gaaggccagt gggtacctgc gggacagctt agcctctaca tcaactgcca gttccattga 3541 caaggccatg cagctgctcc tgtgtgatct acttcttgtg gcccgcacca gcctatggcg 3601 gcgccaacag tcagcagctt cagcccaggg agctcacggt accagcaatg gaccccaggc 3661 ctctgctctg gagctgcgtg gtttccaaca tgacctgagc agcctgaggc gcttggcaca 3721 gagcttccgg cctgctatga ggagggtctt cctacatgag gccacagctc ggctgatggc 3781 aggagcaagt cctgcccgga cacaccagct cctggaccgc agtctgcgga ggcgggcagg 3841 ttccagtggc aaaggaggcg ctgcagctga gctggagcct cgacccacat ggcgggagca 3901 cacagaggcc ttgctgttgg cctcctgcta tctgccccct gccttcctgt cggcccccgg 3961 gcagcgagtg agcatgctgg ctgaggcagc gcgcaccgtg gagaagcttg gcgatcaccg 4021 gctcctgctt gactgccagc agatgctcct gcgcctgggt ggcgggacca ctgtcacttc 4081 cagcctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 4141 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa 4201 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 4261 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 4321 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 4381 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 4441 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 4501 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 4561 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 4621 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 4681 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 4741 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga 4801 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 4861 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 4921 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 4981 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 5041 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 5101 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 5161 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 5221 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc 5281 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 5341 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 5401 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt 5461 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 5521 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 5581 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 5641 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 5701 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 5761 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 5821 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 5881 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 5941 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 6001 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 6061 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 6121 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 6181 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 6241 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 6301 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 6361 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 6421 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 6481 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 6541 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 6601 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 6661 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 6721 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 6781 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 6841 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 6901 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 6961 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 7021 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 7081 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 7141 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 7201 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 7261 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 7321 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 7381 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 7441 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 7501 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 7561 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 7621 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 7681 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 7741 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 7801 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7861 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 7921 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7981 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 8041 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 8101 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 8161 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 8221 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 8281 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 8341 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 8401 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 8461 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 8521 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 8581 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 8641 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 8701 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 8761 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 8821 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 8881 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 8941 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 9001 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 9061 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 9121 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 9181 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg 9241 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 9301 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 9361 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg //