LOCUS V023791 8069 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023791 VERSION V023791 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..3559 /label="VLDLR(NM_003383)" /note="VLDLR(NM_003383)" /gene="VLDLR" CDS 3566..3631 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3669..3893 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3939..4367 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4381..4710 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4777..5571 /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 5745..5866 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5915..5931) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5939..5955 /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(5963..5993) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6008..6029 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6317..6902) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7073..7933) /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(7934..8038) /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 atgggcacgt ccgcgctctg 961 ggcgctctgg ctgctgctcg cgctgtgctg ggcgccccgg gagagcggcg ccaccggaac 1021 cgggagaaaa gccaaatgtg aaccctccca attccagtgc acaaatggtc gctgtattac 1081 gctgttgtgg aaatgtgatg gggatgaaga ctgtgttgac ggcagtgatg aaaagaactg 1141 tgtaaagaag acgtgtgctg aatctgactt cgtgtgcaac aatggccagt gtgttcccag 1201 ccgatggaag tgtgatggag atcctgactg cgaagatggt tcagatgaaa gcccagaaca 1261 gtgccatatg agaacatgcc gcatacatga aatcagctgt ggcgcccatt ctactcagtg 1321 tatcccagtg tcctggagat gtgatggtga aaatgattgt gacagtggag aagatgaaga 1381 aaactgtggc aatataacat gtagtcccga cgagttcacc tgctccagtg gccgctgcat 1441 ctccaggaac tttgtatgca atggccagga tgactgcagc gatggcagtg atgagctgga 1501 ctgtgccccg ccaacctgtg gcgcccatga gttccagtgc agcacctcct cctgcatccc 1561 catcagctgg gtatgcgacg atgatgcaga ctgctccgac caatctgatg agtccctgga 1621 gcagtgtggc cgtcagccag tcatacacac caagtgtcca gccagcgaaa tccagtgcgg 1681 ctctggcgag tgcatccata agaagtggcg atgtgatggg gaccctgact gcaaggatgg 1741 cagtgatgag gtcaactgtc cctctcgaac ttgccgacct gaccaatttg aatgtgagga 1801 tggcagctgc atccatggca gcaggcagtg taatggtatc cgagactgtg tcgatggttc 1861 cgatgaagtc aactgcaaaa atgtcaatca gtgcttgggc cctggaaaat tcaagtgcag 1921 aagtggagaa tgcatagata tcagcaaagt atgtaaccag gagcaggact gcagggactg 1981 gagtgatgag cccctgaaag agtgtcatat aaacgaatgc ttggtaaata atggtggatg 2041 ttctcatatc tgcaaagacc tagttatagg ctacgagtgt gactgtgcag ctgggtttga 2101 actgatagat aggaaaacct gtggagatat tgatgaatgc caaaatccag gaatctgcag 2161 tcaaatttgt atcaacttaa aaggcggtta caagtgtgaa tgtagtcgtg gctatcaaat 2221 ggatcttgct actggcgtgt gcaaggcagt aggcaaagag ccaagtctga tcttcactaa 2281 tcgaagagac atcaggaaga ttggcttaga gaggaaagaa tatatccaac tagttgaaca 2341 gctaagaaac actgtggctc tcgatgctga cattgctgcc cagaaactat tctgggccga 2401 tctaagccaa aaggctatct tcagtgcctc aattgatgac aaggttggta gacatgttaa 2461 aatgatcgac aatgtctata atcctgcagc cattgctgtt gattgggtgt acaagaccat 2521 ctactggact gatgcggctt ctaagactat ttcagtagct accctagatg gaaccaagag 2581 gaagttcctg tttaactctg acttgcgaga gcctgcctcc atagctgtgg acccactgtc 2641 tggctttgtt tactggtcag actggggtga accagctaaa atagaaaaag caggaatgaa 2701 tggattcgat agacgtccac tggtgacagc ggatatccag tggcctaacg gaattacact 2761 tgaccttata aaaagtcgcc tctattggct tgattctaag ttgcacatgt tatccagcgt 2821 ggacttgaat ggccaagatc gtaggatagt actaaagtct ctggagttcc tagctcatcc 2881 tcttgcacta acaatatttg aggatcgtgt ctactggata gatggggaaa atgaagcagt 2941 ctatggtgcc aataaattca ctggatcaga gctagccact ctagtcaaca acctgaatga 3001 tgcccaagac atcattgtct atcatgaact tgtacagcca tcaggtaaaa attggtgtga 3061 agaagacatg gagaatggag gatgtgaata cctatgcctg ccagcaccac agattaatga 3121 tcactctcca aaatatacct gttcctgtcc cagtgggtac aatgtagagg aaaatggccg 3181 agactgtcaa agtactgcaa ctactgtgac ttacagtgag acaaaagata cgaacacaac 3241 agaaatttca gcaactagtg gactagttcc tggagggatc aatgtgacca cagcagtatc 3301 agaggtcagt gttcccccaa aagggacttc tgccgcatgg gccattcttc ctctcttgct 3361 cttagtgatg gcagcagtag gtggctactt gatgtggcgg aattggcaac acaagaacat 3421 gaaaagcatg aactttgaca atcctgtgta cttgaaaacc actgaagagg acctctccat 3481 agacattggt agacacagtg cttctgttgg acacacgtac ccagcaatat cagttgtaag 3541 cacagatgat gatctagctc tcgaggacta caaagaccat gacggtgatt ataaagatca 3601 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc 3661 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 3721 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 3781 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 3841 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 3901 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 3961 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 4021 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 4081 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 4141 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 4201 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 4261 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 4321 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 4381 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 4441 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 4501 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 4561 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 4621 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 4681 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 4741 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 4801 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 4861 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 4921 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 4981 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 5041 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 5101 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 5161 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 5221 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 5281 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 5341 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 5401 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 5461 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 5521 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 5581 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 5641 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 5701 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 5761 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5821 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 5881 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 5941 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 6001 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 6061 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 6121 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 6181 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 6241 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 6301 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 6361 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 6421 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 6481 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 6541 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 6601 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 6661 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 6721 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 6781 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 6841 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 6901 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6961 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 7021 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 7081 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 7141 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 7201 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 7261 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 7321 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 7381 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 7441 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 7501 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 7561 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 7621 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 7681 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 7741 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 7801 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 7861 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 7921 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7981 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 8041 catttccccg aaaagtgcca cctgacgtc //