LOCUS V032311 8629 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032311 VERSION V032311 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 706..3285 /label="LDLR(NM_000527)" /note="LDLR(NM_000527)" /gene="LDLR" CDS 3292..3381 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 3405..3993 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4059..4107 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4309..4737 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4751..5080 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5147..5941 /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 6117..6238 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6287..6303) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6311..6327 /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(6335..6365) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6380..6401 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6689..7277) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7448..8308) /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(8309..8413) /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggg gccctggggc 721 tggaaattgc gctggaccgt cgccttgctc ctcgccgcgg cggggactgc agtgggcgac 781 agatgcgaaa gaaacgagtt ccagtgccaa gacgggaaat gcatctccta caagtgggtc 841 tgcgatggca gcgctgagtg ccaggatggc tctgatgagt cccaggagac gtgcttgtct 901 gtcacctgca aatccgggga cttcagctgt gggggccgtg tcaaccgctg cattcctcag 961 ttctggaggt gcgatggcca agtggactgc gacaacggct cagacgagca aggctgtccc 1021 cccaagacgt gctcccagga cgagtttcgc tgccacgatg ggaagtgcat ctctcggcag 1081 ttcgtctgtg actcagaccg ggactgcttg gacggctcag acgaggcctc ctgcccggtg 1141 ctcacctgtg gtcccgccag cttccagtgc aacagctcca cctgcatccc ccagctgtgg 1201 gcctgcgaca acgaccccga ctgcgaagat ggctcggatg agtggccgca gcgctgtagg 1261 ggtctttacg tgttccaagg ggacagtagc ccctgctcgg ccttcgagtt ccactgccta 1321 agtggcgagt gcatccactc cagctggcgc tgtgatggtg gccccgactg caaggacaaa 1381 tctgacgagg aaaactgcgc tgtggccacc tgtcgccctg acgaattcca gtgctctgat 1441 ggaaactgca tccatggcag ccggcagtgt gaccgggaat atgactgcaa ggacatgagc 1501 gatgaagttg gctgcgttaa tgtgacactc tgcgagggac ccaacaagtt caagtgtcac 1561 agcggcgaat gcatcaccct ggacaaagtc tgcaacatgg ctagagactg ccgggactgg 1621 tcagatgaac ccatcaaaga gtgcgggacc aacgaatgct tggacaacaa cggcggctgt 1681 tcccacgtct gcaatgacct taagatcggc tacgagtgcc tgtgccccga cggcttccag 1741 ctggtggccc agcgaagatg cgaagatatc gatgagtgtc aggatcccga cacctgcagc 1801 cagctctgcg tgaacctgga gggtggctac aagtgccagt gtgaggaagg cttccagctg 1861 gacccccaca cgaaggcctg caaggctgtg ggctccatcg cctacctctt cttcaccaac 1921 cggcacgagg tcaggaagat gacgctggac cggagcgagt acaccagcct catccccaac 1981 ctgaggaacg tggtcgctct ggacacggag gtggccagca atagaatcta ctggtctgac 2041 ctgtcccaga gaatgatctg cagcacccag cttgacagag cccacggcgt ctcttcctat 2101 gacaccgtca tcagcagaga catccaggcc cccgacgggc tggctgtgga ctggatccac 2161 agcaacatct actggaccga ctctgtcctg ggcactgtct ctgttgcgga taccaagggc 2221 gtgaagagga aaacgttatt cagggagaac ggctccaagc caagggccat cgtggtggat 2281 cctgttcatg gcttcatgta ctggactgac tggggaactc ccgccaagat caagaaaggg 2341 ggcctgaatg gtgtggacat ctactcgctg gtgactgaaa acattcagtg gcccaatggc 2401 atcaccctag atctcctcag tggccgcctc tactgggttg actccaaact tcactccatc 2461 tcaagcatcg atgtcaacgg gggcaaccgg aagaccatct tggaggatga aaagaggctg 2521 gcccacccct tctccttggc cgtctttgag gacaaagtat tttggacaga tatcatcaac 2581 gaagccattt tcagtgccaa ccgcctcaca ggttccgatg tcaacttgtt ggctgaaaac 2641 ctactgtccc cagaggatat ggttctcttc cacaacctca cccagccaag aggagtgaac 2701 tggtgtgaga ggaccaccct gagcaatggc ggctgccagt atctgtgcct ccctgccccg 2761 cagatcaacc cccactcgcc caagtttacc tgcgcctgcc cggacggcat gctgctggcc 2821 agggacatga ggagctgcct cacagaggct gaggctgcag tggccaccca ggagacatcc 2881 accgtcaggc taaaggtcag ctccacagcc gtaaggacac agcacacaac cacccgacct 2941 gttcccgaca cctcccggct gcctggggcc acccctgggc tcaccacggt ggagatagtg 3001 acaatgtctc accaagctct gggcgacgtt gctggcagag gaaatgagaa gaagcccagt 3061 agcgtgaggg ctctgtccat tgtcctcccc atcgtgctcc tcgtcttcct ttgcctgggg 3121 gtcttccttc tatggaagaa ctggcggctt aagaacatca acagcatcaa ctttgacaac 3181 cccgtctatc agaagaccac agaggatgag gtccacattt gccacaacca ggacggctac 3241 agctacccct cgagacagat ggtcagtctg gaggatgacg tggcgctcga ggaacaaaaa 3301 ctaatatcag aggaagattt ggaacagaaa ctaatcagtg aagaagattt agagcaaaag 3361 ttaatttccg aggaggactt atagtgaggg cccgatatct cgacaatcaa cctctggatt 3421 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 3481 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 3541 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 3601 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 3661 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 3721 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 3781 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 3841 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 3901 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 3961 cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac 4021 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa 4081 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact 4141 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc 4201 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag 4261 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta 4321 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 4381 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 4441 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc 4501 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 4561 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 4621 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg 4681 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat 4741 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 4801 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 4861 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 4921 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 4981 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa 5041 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat 5101 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 5161 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 5221 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 5281 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc 5341 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 5401 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 5461 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 5521 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 5581 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 5641 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt 5701 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 5761 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 5821 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 5881 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 5941 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 6001 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 6061 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 6121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 6181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 6241 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 6301 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 6361 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 6421 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 6481 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 6541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 6601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 6661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 6721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 6781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 6841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt 6901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 6961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 7021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 7081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg 7141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 7201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 7261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 7321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 7381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 7441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 7501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 7561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 7621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 7681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 7741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 7801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 7861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 7921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 7981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 8041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 8101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 8161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 8221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 8281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 8341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 8401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc 8461 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca 8521 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc 8581 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg //